Improved estimates of abortion rates in tsetse (Glossina spp.).

Abortion rates were assessed among 170, 846 tsetse (154,228 Glossina pallidipes and 19,618 Glossina morsitans morsitans) sampled in Zimbabwe in 1988-1999. The study produced improved estimates of abortion rates and how these varied with fly age and size and temperatures experienced during pregnancy. An abortion was diagnosed if the uterus was empty and the largest oocyte <0.82 of the expected mature length. Abortion rates for G. pallidipes and G. m. morsitans were 0.64% (95% ci: 0.59-0.69) and 0.83% (0.62-1.10) for trapped flies and 2.03% (1.77-2.31) and 1.55% (1.20-1.98) for flies from artificial refuges. Abortion rates increased with increasing temperature and decreased with increasing wing length and wing fray. Contrary to laboratory findings, abortion rates did not increase in the oldest flies. Percentages of tsetse with empty uteri, regardless of abortion status, were significantly higher than estimated abortion percentages. For tsetse from traps, 4.01% (95% ci: 3.90-4.13) of G. pallidipes and 2.52% (2.14-2.95) of G. m. morsitans had empty uteri; for flies from artificial refuges, the percentages were 12.69% (12.07-13.34) and 14.90% (13.82-16.02), respectively. Abortion losses are small relative to losses at all other stages of life.

Improved models for the relationship between age and the probability of trypanosome infection in female tsetse, Glossina pallidipes Austen.

Between 1990 and 1999, at Rekomitjie Research Station, Zambezi Valley, Zimbabwe, 29,360 female G. pallidipes were dissected to determine their ovarian category and trypanosome infection status. Overall prevalences were 3.45 and 2.66% for T. vivax and T. congolense, respectively, declining during each year as temperatures increased from July - December. Fits to age-prevalence data using Susceptible-Exposed-Infective (SEI) and SI compartmental models were statistically better than those obtained using a published catalytic model, which made the unrealistic assumption that no female tsetse survived more than seven ovulations. The improved models require knowledge of fly mortality, estimated separately from ovarian category distributions. Infection rates were not significantly higher for T. vivax than for T. congolense. For T. congolense in field-sampled female G. pallidipes, we found no statistical support for a model where the force of infection was higher at the first feed than subsequently. The long survival of adult female tsetse, combined with feeding at intervals ≤3 days, ensures that post-teneral feeds, rather than the first feed, play the dominant role in the epidemiology of T. congolense infections in G. pallidipes. This is supported by estimates that only about 3% of wild hosts at Rekomitjie were harbouring sufficient T. congolense to ensure that tsetse feeding off them take an infected meal, so that the probability of ingesting an infected meal is low at every meal.

Negative density-dependent dispersal in tsetse (Glossina spp): red flag or red herring?

A deterministic model of the distribution of tsetse flies (Glossina spp) was used to assess the extent to which the efficacy of control operations would be affected by three different modes of density dependence in per capita adult dispersal: (i) density-independent dispersal which has been commonly adopted in previous models, (ii) positive density-dependent dispersal which has occasionally been discussed in the tsetse literature, (iii) negative density-dependent dispersal (NDDD). The last has recently been suggested, from genetic studies, to change the dispersal rate of tsetse by up to 200-fold, thereby posing a severe risk for the success of tsetse control operations. Modelling outputs showed that NDDD poses no such risk, provided the mean daily dispersal of tsetse is below about 1 km, which is greater than any rate actually recorded in the field or indicated by the genetic studies. NDDD can be problematic only if tsetse disperse at rates that appear highly unlikely, or even impossible, on energetic grounds. Under some circumstances these high rates would help rather than hinder the control officer. NDDD is not necessary to explain the results of control operations, and not sufficient to explain the results of successful control programmes.

A model for the relationship between wing fray and chronological and ovarian ages in tsetse (Glossina spp).

Age-dependent mortality changes in haematophagous insects are difficult to measure but are important determinants of population dynamics and vectorial capacity. A Markov process was used to model age-dependent changes in wing fray in tsetse (Glossina spp), calibrated using published mark-recapture data for male G. m. morsitans in Tanzania. The model was applied to female G. m. morsitans, captured in Zimbabwe using a vehicle-mounted electric net and subjected to ovarian dissection and wing fray analysis. Rates of fray increased significantly with age in males but not females, where the rate was constant for ovarian categories 0-3. A jump in mean fray between ovarian categories 3 and 4 + 4n is consistent with the latter category including flies that have ovulated 4, 8, 12, 16 times and so on. The magnitude of the jump could, theoretically, facilitate improved mortality estimates. In practice, although knowledge of fly mortality was required for modelling wing fray, mortality estimates derived from ovarian dissection data are independent of patterns and rates of change in wing fray. Significantly better fits to ovarian age data resulted when age-specific mortality was modelled as the sum of two exponentials, with high mortality in young and old flies, than when mortality was constant at 2.3% per day.

Estimating tsetse fertility: daily averaging versus periodic larviposition.

When computing mean daily fertility in adult female tsetse, the common practice of taking the reciprocal of the interlarval period (called averaged fertility) was compared with the method of taking the sum of the products of daily fertility and adult survivorship divided by the sum of daily survivorships (called periodic fertility). The latter method yielded a consistently higher measure of fertility (approximately 10% for tsetse) than the former method. A conversion factor was calculated to convert averaged fertility to periodic fertility. A feasibility criterion was determined for the viability of a tsetse population. Fertility and survivorship data from tsetse populations on Antelope Is. and Redcliff Is., both in Zimbabwe, were used to illustrate the feasibility criterion, as well as the limitations imposed by survivorship and fertility on the viability of tsetse populations. The 10% difference in fertility between the two methods of calculation makes the computation of population feasibility with some parameter combinations sometimes result in a wrong answer. It also underestimates both sterile male release rates required to eradicate a pest population, as well as the speed of resurgence if an eradication attempt fails.

Modelling optimal timing and frequency of insecticide sprays for eradication or knockdown of closed populations of tsetse flies Glossina spp. (Diptera: Glossinidae).

A population model for tsetse species was used to assess the optimal number and spacing of airborne sprays to reduce or eradicate a tsetse population. It was found that the optimal spray spacing was determined by the time (days) from adult emergence to the first larviposition and, for safety, spacing was assigned to that duration minus 2 days. If sprays killed all adults, then the number of sprays required for eradication is determined by a simple formula. If spray efficiency is less than 100% kill per spray, then a simulation was used to determine the optimal number, which was strongly affected by spray efficiency, mean daily temperature, pupal duration, age to first larviposition and the acceptance threshold for control, rather than eradication. For eradication, it is necessary to have a spray efficiency of greater than 99.9% to avoid requiring an excessive number of sprays. Output from the simulation was compared with the results of two aerial spraying campaigns against tsetse and a least squares analysis estimated that, in both cases, the kill efficiency of the sprays was not significantly less than 100%.

Models for the rates of pupal development, fat consumption and mortality in tsetse (Glossina spp).

Environmental temperature is an important driver of the population dynamics of tsetse (Glossina spp) because the fly's immature stages are particularly vulnerable to temperatures (T) outside the range T = 16-32°C. Laboratory experiments carried out 50 years ago provide extensive measures of temperature-dependent rates of development, fat consumption and mortality in tsetse pupae. We improve on the models originally fitted to these data, providing better parameter estimates for use in population modelling. A composite function accurately models rates of pupal development for T = 8-32°C. Pupal duration can be estimated by summing the temperature-dependent daily percentage of development completed. Fat consumption is modelled as a logistic function of temperature; the total fat consumed during pupal development takes a minimum for T ≈ 25°C. Pupae experiencing constant temperatures <16°C exhaust their fat reserves before they complete development. At high temperatures, direct effects kill the pupae before fat stores are exhausted. The relationship between pupal mortality and temperature is well described by the sum of two exponential functions. Summing daily mortality rates over the whole pupal period does not reliably predict overall mortality. Mortality is more strongly correlated with the mean temperature experienced over pupal life or, for T ≤ 30°C, the fat consumption during this period. The new results will be particularly useful in the construction of various models for tsetse population dynamics, and will have particular relevance for agent-based models where the lives of individual tsetse are simulated using a daily time step.

Is Zimbabwe ready to transition from anonymous unlinked sero-surveillance to using prevention of mother to child transmission of HIV (PMTCT) program data for HIV surveillance?: results of PMTCT utility study, 2012.

BACKGROUND: Prevention of mother-to-child transmission of HIV (PMTCT) programs collect socio-demographic and HIV testing information similar to that collected by unlinked anonymous testing sero-surveillance (UAT) in antenatal settings. Zimbabwe evaluated the utility of PMTCT data in replacing UAT. METHODS: A UAT dataset was created by capturing socio-demographic, testing practices from the woman's booking-card and testing remnant blood at a laboratory from 1 June to 30 September 2012. PMTCT data were collected retrospectively from ANC registers. UAT and PMTCT data were linked by bar-code labels that were temporarily affixed to the ANC register. A questionnaire was used to obtain facility-level data at 53 sites. RESULTS: Pooled HIV prevalence was 15.8 % (95 % CI 15.3-16.4) among 17,349 women sampled by UAT, and 16.3 % (95 % CI 15.8 %-16.9 %) among 17,150 women in PMTCT datasets for 53 sites. Pooled national percent-positive agreement (PPA) was 91.2 %, and percent-negative agreement (PNA) was 98.7 % for 16,782 women with matched UAT and PMTCT data. Based on UAT methods, overall median prevalence was 12.9 % (Range 4.0 %-19.4 %) among acceptors and refusers of HIV test in PMTCT compared to 12.5 % ((Range 3.4 %-19.5 %) among acceptors in ANC registers. There were variations in prevalence by site. CONCLUSION: Although, there is no statistical difference between pooled HIV prevalence in UAT compared to PMTCT program, the overall PPA of 91.2 % and PNA of 98.7 % fall below World Health Organisation (WHO) benchmarks of 97.6 % and 99.6 % respectively. Zimbabwe will need to strengthen quality assurance (QA) of rapid HIV testing and data collection practices. Sites with good performance should be prioritised for transitioning.

Mortality estimates from ovarian age distributions of the tsetse fly Glossina pallidipes Austen sampled in Zimbabwe suggest the need for new analytical approaches.

Mortality estimates are central to understanding tsetse fly population dynamics, but are difficult to acquire from wild populations. They can be obtained from age distribution data but, with limited data, it is unclear whether the assumptions required to make the estimates are satisfied and, if not, how violations affect the estimates. We evaluate the assumptions required for existing mortality estimation techniques using long-term longitudinal ovarian dissection data from 144,106 female tsetse, Glossina pallidipes Austen, captured in Zimbabwe between 1988 and 1999. At the end of the hot-dry season each year, mean ovarian ages peaked, and maximum-likelihood mortality estimates declined to low levels, contrary to mark-recapture estimates, suggesting violations of the assumptions underlying the estimation technique. We demonstrate that age distributions are seldom stable for G. pallidipes at our study site, and hypothesize that this is a consequence of a disproportionate increase in the mortality of pupae and young adults at the hottest times of the year. Assumptions of age-independent mortality and capture probability are also violated, the latter bias varying with capture method and with pregnancy and nutritional status. As a consequence, mortality estimates obtained from ovarian dissection data are unreliable. To overcome these problems we suggest simulating female tsetse populations, using dynamical modelling techniques that make no assumptions about the stability of the age distribution.

A general model for mortality in adult tsetse (Glossina spp.).

Tsetse exhibit a U-shaped age-mortality curve, with high losses after eclosion and a well-marked ageing process, which is particularly dramatic in males. A three-parameter (k(1) -k(3) ) model for age-dependent adult instantaneous mortality rates was constructed using mark-recapture data for the tsetse fly Glossina morsitans morsitans Westwood (Diptera: Glossinidae). Mortality changed linearly with k(1) over all ages; k(2) affected only losses in roughly the first week of adult life, and k(3) controlled the ageing rate. Mortality pooled over age was twice as sensitive to changes in k(3) as in k(1) . Population growth rate was, however, similarly affected by these two parameters, reflecting the disproportionate effect of k(3) on mortality in the oldest flies that contribute least to the growth rate. Pooled-age mortality and growth rate were insensitive to changes in k(2) . The same model also provided good fits to data for laboratory colonies of female G. m. morsitans and Glossina austeni Newstead and should be applicable to all tsetse of both sexes. The new model for tsetse mortality should be incorporated into models of tsetse and trypanosome population dynamics; it will also inform the estimation of adult female mortality from ovarian dissection data.

A randomised placebo-controlled trial to explore the effect of suppressive therapy with acyclovir on genital shedding of HIV-1 and herpes simplex virus type 2 among Zimbabwean sex workers.

OBJECTIVES: To determine the effect of daily acyclovir on genital shedding of HIV-1 and herpes simplex virus type 2 (HSV-2) in a randomised placebo-controlled trial among rural Zimbabwean sex workers. METHODS: 214 women were recruited and tested for HIV-1 and HSV-2 antibodies, HIV plasma viral load, CD4 lymphocyte count and genital swabs for qualitative detection of HIV-1 and HSV-2 genital shedding. Women were randomly assigned to acyclovir 400 mg twice a day for 12 weeks or matching placebo and were followed weekly to detect HIV-1 or HSV-2 genital shedding. Shedding analyses were only undertaken on 125 women co-infected with HSV-2 and HIV-1. Data were analysed using logistic regression, with random effects modelling used to account for repeated measurements on the same women. RESULTS: All women were randomly assigned to acyclovir or placebo; 125 of whom were co-infected with HIV-1 and HSV-2. 69 women were randomly assigned to acyclovir and 56 to placebo. Although twice daily acyclovir reduced rates of HSV-2 genital shedding, (adjusted odds ratio (AOR) 0.24; 95% CI 0.12 to 0.48; less than p<0.001), it had no effect on the proportion of visits at which HIV-1 shedding was detected (AOR 1.08; 95% CI 0.48 to 2.42; p = 0.9). Adherence varied between participants but even when adherence was high (as determined by pill count and extent of HSV-2 suppression) HIV-1 shedding was not reduced. CONCLUSION: Among these HIV-1 and HSV-2-seropositive women, suppressive acyclovir therapy had no effect on the rate of HIV genital shedding despite a reduction in genital HSV-2. Treatment adherence and its measurement clearly affect the interpretation of these results.

Enhancement of resistance to coccidiosis and necrotic enteritis in broiler chickens by dietary muscadine pomace.

Muscadine pomace (MP), a by-product of the production of wine and juice from Vitis rotundifolia, was dried and tested in chickens for effects on primary resistance to coccidiosis, development of protective immunity after vaccination with live coccidia, and resistance to necrotic enteritis (NE) caused by the joint action of Clostridium perfringens and coccidia. To test primary resistance to coccidiosis, 2-wk-old chicks were given 2% or 5% MP in the diet and inoculated with Eimeria acervulina and E. maxima. Birds given MP at either level had significantly (P < 0.05) lower lesion scores at 7 days postinoculation, in comparison with control birds, although weight gains were statistically similar. Broiler chickens were given 2% or 5% MP and grown to 42 days to test the palatability of MP. Birds given 2% MP in feed grew similarly to untreated controls, but birds given 5% had poorer average live weight. This suggested a negative effect on feed intake at the higher level. The effects of dietary 0.5% or 2.0% MP on immune protection were tested after live coccidiosis vaccination in the hatchery. Chicks were removed from each pen at 21 days of age and challenged with E acervulina, E. maxima, and E. tenella. Resistance to infection was improved by MP as suggested by significantly (P < 0.05) lower lesion scores 7 days postchallenge, and improved weight gains in comparison with immunized control birds that did not receive MP. At 42 days of age, birds given MP had higher average live weights than controls, although feed efficiency was not affected. An established model was used to study the effect of MP on NE in broiler chickens. Chicks were inoculated with live coccidia at 14 days of age and dosed orally with live cultures of C perfringens on day 19, day 20, and day 21. Enteritis caused 48% mortality in the first study and 67% mortality in the second study. Dietary MP at 0.5-2.0% significantly (P < 0.05) reduced mortality in both experiments; improved weight gain relative to the unmedicated, infected control; and reduced lesion scores at necropsy. Overall, the results of six experiments suggested that MP given in the diet at 0.5% or higher had a positive effect on primary resistance and development of acquired resistance to two severe intestinal diseases in chickens.

Comparison of specimens for detection of porcine reproductive and respiratory syndrome virus infection in boar studs.

Porcine reproductive and respiratory syndrome virus (PRRSV)-contaminated semen from boars is a route of transmission to females, and early detection of PRRSV infection in boars is a key component in sow farm biosecurity. The purpose of this study was to determine the optimum diagnostic specimen(s) for the detection of acute PRRSV infection in boars. Individually housed boars (n = 15) were trained for semen and oral fluid collection and then vaccinated with a commercial PRRSV modified live virus vaccine. Starting on the day of vaccination and for 14 days thereafter, oral fluid specimens were collected daily from all boars. The 15 boars were subdivided into three groups of 5, and serum, blood swabs and 'frothy saliva' were collected at the time of semen collection on a 3-day rotation. Frothy saliva, derived from the submandibular salivary gland, is produced by aroused boars. Semen was centrifuged, and semen supernatant and cell fractions were tested separately. All samples were randomly ordered and then tested by PRRSV real-time quantitative reverse-transcription polymerase chain reaction assay (rRT-PCR) and PRRSV antibody ELISA. In this study, a comparison of serum, blood swab, and oral fluid rRT-PCR results found no statistically significant differences in the onset of detection or proportion of positives, but serum was numerically superior to oral fluids for early detection. Serum and oral fluid provided identical rRT-PCR results at ≥ 5 day post-vaccination. Likewise, the onset of detection of PRRSV antibody in serum, oral fluid and frothy saliva was statistically equivalent, with serum results again showing a numerical advantage. These results showed that the highest assurance of providing PRRSV-negative semen to sow farms should be based on rRT-PCR testing of serum collected at the time of semen collection. This approach can be augmented with oral fluid sampling from a random selection of uncollected boars to provide for statistically valid surveillance of the boar stud.

Degradation of tyrosine aminotransferase (TAT) via the ubiquitin-proteasome pathway.

Most of the known cellular substrates of the ubiquitin system are short-lived growth regulators and transcriptional activators. Very few enzymes involved in intermediary metabolism have been shown to be targeted by the system. In a reconstituted cell-free system, we show that tyrosine aminotransferase (TAT), a key enzyme involved in amino acid metabolism, is conjugated and degraded in an ATP- and ubiquitin-dependent manner. Degradation of ubiquitin-TAT adducts requires, in addition to the 26S proteasome, a novel, yet unidentified, factor. TAT can be protected from degradation by association with its coenzyme pyridoxal phosphate. To examine the potential role of the ubiquitin system in regulating the stability of the enzyme in vivo, we show that cell extracts derived from livers of animals in which TAT was induced, display a corollary increase in the formation of specific TAT-ubiquitin adducts.

From genotype to phenotype: computer-based modeling of gene expression with STELLA II.

STELLA II is a microcomputer program that is designed to simulate and animate the dynamics generated by biological systems in a quantitative fashion. The program employs the Macintosh user interface and includes a tool kit for assembling models, plus menus for setting parameters and selecting input or output. Quantitative relationships among components may be expressed with user-defined equations, graphical functions or data from spreadsheets. System dynamics are simulated by running the program after defining initial conditions, and results may be viewed immediately using windows for graphical or tabular plots. The present article describes how STELLA II may be used to simulate gene expression as an adjunct to experimentation or student-directed learning.

A computer program for modeling the kinetics of gene expression.

Enzyme induction may be modeled on the basis of four, quantifiable processes that control the rates at which specific gene products accumulate and decay. These processes include synthesis of functional mRNA, translation and degradation of mRNA, and degradation of the protein product. We present a simple computer program that permits mathematical simulation of gene expression on the basis of experimentally determined rates of synthesis and degradation. The program was implemented as a spreadsheet using Microsoft Excel for Macintosh and MS-DOS operating systems and also was adapted for HyperCard on the Macintosh. It contains a formula to account for growth of tissue or cell populations. The program predicts amounts of individual mRNAs and proteins (or enzyme activities) in cells as a function of time after a stimulus alters their rates of synthesis or degradation.

Experimental diabetes increases the formation of sulfane by transsulfuration and inactivation of tyrosine aminotransferase in cytosols from rat liver.

The addition of L-cysteine to hepatic cytosols causes inactivation of tyrosine aminotransferase. We have studied the mechanism of inactivation and the effect of streptozotocin-induced diabetes in the rat on the inactivation of tyrosine aminotransferase in the presence of fractions prepared from livers and kidneys. Diabetes increased the rate at which tyrosine aminotransferase was inactivated after addition of cysteine to hepatic cytosols. The inactivation was due to the production of thiocysteine (which contains sulfane sulfur) from cystine as a result of desulfuration catalyzed by gamma-cystathionase. Diabetes increased the content of cystathionine beta-synthase and gamma-cystathionase in liver. As a result, cytosols from diabetic animals converted homocysteine, cystathionine, cysteine and cystine to sulfane at an elevated rate, with resulting inactivation of tyrosine aminotransferase. In contrast, inactivation in kidney fractions was not affected by diabetes. Incubation with an inhibitor of gamma-cystathionase (propargylglycine) prevented inactivation of tyrosine aminotransferase. These results show that the potential for the formation of sulfane sulfur by the enzymes of the transsulfuration pathway is enhanced by chronic diabetes.

Maternal pulmonary edema during premature labor inhibition.

Three cases of pulmonary edema after treatment with terbutaline are reported. In 1 patient invasive hemodynamic monitoring shortly after the onset of pulmonary edema showed normal pressures and pulmonary vascular resistance with augmented systemic blood flow. These findings suggest that this syndrome might arise on a noncardiogenic basis.

Menopausal hormone replacement therapy with continuous daily oral micronized estradiol and progesterone.

The safety and efficacy of a daily combination of micronized estradiol (E2) (0.7-1.05 mg) and progesterone (200-300 mg) were evaluated in ten menopausal women with moderate to severe vasomotor symptoms and/or vaginal atrophy over a 12-month study interval. For comparison, five similar women were placed on conjugated estrogens, 0.625 mg daily, and medroxyprogesterone acetate, 10 mg daily, for the first 10 days of each calendar month for 12 months. Patients were evaluated at 0, 1, 3, 6, and 12 months. Estrogens rose significantly from baseline in both groups (P less than .01). Progesterone increased significantly above baseline in the E2 and progesterone group (P less than .01), but did not change in the conjugated estrogens and medroxyprogesterone acetate users. All women on E2 and progesterone had a decrease in total cholesterol and an increase in high-density lipoprotein cholesterol from baseline (P less than .01). Those on conjugated estrogens and medroxyprogesterone acetate had no significant change from baseline in total cholesterol; however, they did have an increase in high-density lipoprotein cholesterol values (P less than .01). In the E2 and progesterone group, the endometrial histology became completely quiescent and there was no uterine bleeding after 6 months of observation. Four of five women on conjugated estrogens and medroxyprogesterone acetate continued regular withdrawal bleeding throughout the study period, but no endometrial hyperplasia was encountered. This study demonstrates that the daily administration of a combination of micronized E2 and progesterone results in symptomatic improvement, minimal side effects, an improved lipid profile, and amenorrhea without endometrial proliferation or hyperplasia in menopausal women.

Differences in glycogen, lipids, and enzymes in livers from rats flown on COSMOS 2044.

Livers from rats flown aboard COSMOS 2044 were analyzed for protein, carbohydrate (glycogen), and lipids as well as the activities of a number of key enzymes involved in metabolism of these compounds and xenobiotics. The major differences between the flight group and the synchronous control were elevations in microsomal protein, liver glycogen content, tyrosine aminotransferase, and tryptophan oxygenase and reductions in sphingolipids and the rate-limiting enzyme of heme biosynthesis, delta-aminolevulinic acid synthase. These results provide further evidence that spaceflight has pronounced and diverse effects on liver function; however, some of the results with samples from COSMOS 2044 differed notably from those from previous spaceflights. This may be due to conditions of spaceflight and/or the postflight recovery period for COSMOS 2044.