Nutritional levels of female tsetse Glossina pallidipes from artificial refuges

Female tsetse flies, Glossina pallidipes Austen, caught in artificial refuges were subjected to ovarian dissection and analysed for levels of fat, residual dry weight (RDW) and haematin. There were rather small proportions of flies in ovarian categories 0 and 1, in part due to large losses in the immature and teneral stages at the hottest time of year. The distribution of the female catch among pregnancy days was close to uniform. The wet and dry weights (WW and DW) and RDW of eggs, larvae and pupae increased by 0.821, 0.303 and 0.204 mg respectively, with each mm3 increase in volume. Water accounted for 71.7% of the fat-free WW, and fat for 32.7% of the DW. Between birth and ovulation, fat increased from 2 to 4 mg and RDW from 7 to 11 mg; thoracic RDW increased by 2.5mg and changed little thereafter. Fat levels increased 3.5mg by day 6 of pregnancy, but only 0.5 mg thereafter. Over the same periods RDW corrected to zero haematin (CRDW) increased by 1 and 8 mg respectively. Full-term fat and CRDW levels were 8.2 and 19.4 mg respectively. Cumulative haematin frequencies formed a smooth curve with a slope that increased continuously. The raw data were well fitted by a model where feeding rates increased exponentially and capture probability was independent of haematin content. The mean feeding interval was 60 h; feeding probabilities of >0.9/day were only found in flies that had failed to feed for>72 h. In early pregnancy, fat levels declined with haematin for flies that had fed>36 h previously; by days 5-7 fat levels were maintained at a constant high level for 60 h post-feeding. Fat-haematin graphs for female tsetse cannot be used to estimate rates of fat utilization. Traps sample tsetse with below-average fat and RDW in early and late pregnancy respectively. Refuge samples are less biased than those from traps; they give a better picture of the dynamics of pregnancy in normal flies and facilitate the explanation of existing anomalies.     

The feeding behaviour, activity and trappability of wild female Glossina pallidipe in relation to their pregnancy cycle

Female Glossina pallidipes Austen trapped with baited NG2B traps were subjected both to detailed ovarian dissection and to nutritional analysis. Using a calibration curve derived from dissected wild-caught, laboratory-held flies, the field females were assigned by discriminant analysis to each day of the pregnancy cycle. Field females were most available to NG2B traps while carrying the first instar larva. The nutritional characteristics of trapped field females over the pregnancy cycle lead to the following main conclusions. (i) Fat levels increase most rapidly during the egg in utero stage, while CRDW increases significantly only during the larval stages, culminating in a 4 mg increase during the last day of the third larval instar. (ii) The haematin content of the flies indicates that females feed at approximately 3-day intervals and may feed on any day of the pregnancy cycle. (iii) The estimated time of feeding during the day corresponds with the observed time of peak activity, both of which are earlier in the day later in the pregnancy cycle. (iv) The rate of fat usage reveals significantly greater flight activity on day 5 of the cycle than on other days, agreeing with the high trappability on this day, and overall females appear to use fat at twice the rate of males. Reproductive data provide a much more accurate picture of the relative sampling efficiency than do nutritional data, although the latter reveal the general trends correctly.     

Nutritional levels of pregnant and postpartum tsetse Glossina pallidipes Austen captured in artificial warthog burrows in the Zambezi Valley of Zimbabwe

In Zimbabwe, female tsetse Glossina pallidipes Austen, collected from artificial warthog burrows and subjected to ovarian dissection and nutritional analysis, provide the first field estimates of resource allocation from mother to offspring across all of pregnancy. Approximately 45% of 1833 females captured are full‐term pregnant on entering the burrow. The remainder presumably use burrows as refuges at temperatures >32 °C. Maternal residual dry weight (RDW) increases by 1.5 mg after the first feed post‐larviposition but, thereafter, only by 1.4 mg by 90% of pregnancy completion. Uterine RDW changes little by 60% of pregnancy completion but increases by >6 mg by parturition. Between the times of 5% and 90% pregnancy completion, maternal RDW is approximately constant: it then declines 2.8 mg by parturition, balancing larval gains of 2.6 mg. Mothers accumulate 6.3 mg of fat in the first 80% of pregnancy, while uterine fat increases by only 0.8 mg. Thereafter, by parturition, larval fat increases by 4.1 mg, whereas maternal fat declines by 3.3 mg. The larva deposited is 5% heavier than its mother and has 52% more fat. RDW and fat levels, corrected for fly size and haematin, are 1 and 2 mg, respectively, lower in females from traps than those from burrows. Burrow catches provide an improved picture of tsetse pregnancy dynamics, highlighting a reproductive strategy involving resource commitment to the larva being delayed until late pregnancy when sufficient stores guarantee viable pupal production. Larval development in tsetse starts significantly later than the analogous changes during pregnancy in two mammals and two viviparous fish.     

A model for blood meal digestion and fat metabolism in male tsetse flies (Glossinidae)

Abstract. Fat and haematin levels of mature male Glossina morsitans morsitans Westwood were estimated at different times after feeding at temperatures between 15 and 30°C. Flies were kept (largely inactive) in 7.5 × 2.5 cm tubes, or in actograph cages, where flight activity increased with time after feeding. Haematin excretion was modelled as a series of three first order reactions, all with the same rate parameter. The model accounted for > 98% of the variance in mean haematin in each of seven experiments; the rate parameter increased linearly with temperature and activity level. A similar approach was adopted for modelling fat metabolism. The rate coefficients of lipogenesis increased with temperature, and that for lipolysis with temperature, activity level and their interaction. All experiments were analysed simultaneously to provide equations predicting haematin or fat levels for all times, for active or inactive flies, and for temperatures between 15 and 30°C. Haematin exhibited large variations between individuals, but for active flies the expected haematin content at a given time varied little between flies kept at 25 and at 30°C. In inactive flies kept at 25°C, lipogenesis peaked at ≈ 24 h and lipolysis at ≈ 48 h. For active flies the times were 12 and 24 h, respectively; both rates were about twice as high as in inactive flies. Active flies produced (up to 1 mg) more fat out of a given size of blood meal than inactive flies. Curves of fat content against logarithm of haematin content differed little with temperature, and can therefore be useful for comparative studies of field populations of tsetse.     

Effect of host odours on trap catch composition of Glossina pallidipes in Kenya

The effect of odour attractants on the composition of samples of Glossina pallidipes Austen was investigated by comparing the age and nutritional status of flies caught in unbaited biconical traps with those caught in traps baited with cow urine and acetone. For both male and female flies, baited traps caught more flies with significantly higher fat content than did unbaited traps. Thus the samples from baited traps were more representative of the population as a whole: males showed a fuller range of the fat/haematin conditions known to occur in the field and proportionately more females were in later stages of the pregnancy cycle, than from unbaited traps.     

The nutritional state of male tsetse flies,Glossina pallidipes,at the time of feeding

Abstract. The feeding intervals of tsetse flies have been estimated from the nutritional state of flies caught in traps. However, such estimates have been disputed on the grounds that traps catch a biased, hungry sample of the flies which are seeking hosts and will feed. In this paper we present data on the nutritional state of tsetse flies caught approaching and feeding on oxen. Individual oxen were surrounded with an incomplete ring of electric nets which caught Glossina pallidipes Austen that were approaching, departing unfed and departing fed from an ox. Non-teneral males caught in this way were analysed for their fat and haematin contents. The feeding interval was estimated from a comparison of the frequency distributions of the pre- and post-feed haematin contents of the flies which fed. The former was not measured directly, and was deduced from the frequency distributions of the haematin contents of the male flies caught approaching and departing unfed from the oxen, since it is assumed that the departing unfed and fed flies together form a sample of the approaching flies. There was no difference between the frequency distributions of haematin contents of flies caught approaching and departing unfed, and therefore the pre-feed haematin contents of the males which fed should have the same frequency distribution. Comparison of this distribution with that of the post-feed haematin contents of the males which fed indicated that the majority of male G.pallidipes were returning to feed after digesting on average 1.4 log haematin units of the previous bloodmeal. From data published elsewhere, this corresponds to a mean feeding interval of 42-60h. There was a strong, linear, negative relationship between the fat contents of males and their probability of taking a bloodmeal, suggesting that fat content is important in determining the feeding behaviour of tsetse flies.     

Physiological correlates of the availability of Glossina morsitans centralis Machado to different sampling methods

Abstract.

• 1 The feeding cycle for male tsetse flies and the pregnancy cycle for females are used as frameworks for investigating the field behaviour of Glossina morsitans centralis Machado in Zambia, and hence the sampling biases of different capture devices.
• 2 Flies were collected in the field using hand nets and an electric back pack on foot patrols, and hand nets, hand-catching and an electric screen on landrover patrols. They were analysed for wing-fray, vein-length, chloroform-extractable fat and haematin. In addition, samples of laboratory-bred females killed daily during their second pregnancy cycle were analysed for the last two parameters.
• 3 The daily flying time of males is estimated from the fat—haematin curve to be about 32 min/day, which is identical to previous estimates for G. morsitans from Tanzania.
• 4 From the varying numbers and fat content of male flies caught by the different sampling methods during the course of the feeding cycle it is concluded that both haematin levels and fat reserves influence fly behaviour.
• 5 Comparison of the frequency distribution of wild-caught and laboratory-bred females along the corrected residual dry weight axis of the pregnancy cycle suggests that the results of combined electric-trap and hand-net sampling reflect the actual frequency distribution in the wild population at least for the first 7 days of the pregnancy cycle; but females in the last 2 days of their pregnancy cycle are apparently unavailable to sampling devices, perhaps because of their reduced flight activity.
• 6 The very variable percentage female catch (10.9–43.4%) returned by the different sampling methods is discussed in the light of the detailed analysis of the differential availability of male and female flies to each capture device.

     

The use of discriminant analysis for the estimation of sampling biases for female tsetse

ABSTRACT.

• 1 In this paper we investigate whether the technique of discriminant analysis can be used to estimate sampling biases for female tsetse.
• 2 Discriminant analysis was first applied to laboratory samples of female tsetse, Glossina morshans morsitans Westwood, to test whether flies of known history could be assigned to the correct day of the pregnancy cycle on the basis of their fat, haematin and corrected residual dry weight.
• 3 Following the satisfactory results from the laboratory samples, the same technique was applied to field samples of G.m. centralis Machado captured by electric traps and hand nets in Zambia and of G.palpalis palpalis (Robineau-Desvoidy) captured in biconical traps at five sites in Ivory Coast. The results show that flies on day 1 of the pregnancy cycle were most likely to be caught, with a second peak of day-6 and day-7 flies, while very few day-8 or day-9 flies were caught.
• 4 These major peaks in fly trappability coincide with the known feeding habits of female tsetse, and indicate synchrony of feeding by many members of the population immediately after larviposition and again as the larva in utero moults from the second to third instar. G.palpalis is relatively more available at this later stage of its pregnancy cycle to the capture methods used than is G.morsitans. A third feed may be taken at a more variable point in the pregnancy cycle.
• 5 This method of estimating the sampling biases of female tsetse could allow an estimate of total population size, as long as the absolute sampling efficiency of flies on any one day of the pregnancy cycle could be established by, for example, mark-release-recapture experiments.

     

Quantitative morphological changes of the uterine gland cells in relation to physiological events during a pregnancy cycle in Glossina morsitans morsitans

Secretory cells of the uterine gland were investigated morphometrically during the second pregnancy cycle in Glossina morsitans morsitans. In early pregnancy when an egg was present in the uterus, morphometric parameters of the nucleus, the cytoplasm and its organelles were markedly low coincident with low physiological activity in the gland cells. This is the period when a proportion of the assimilated amino acids from the digested blood meals are converted to triglyceride in the fat body and stored. During the second half of pregnancy, when a larva was present in the uterus, there was a striking increase of the cytoplasmic and organelle volumes as well as of the surface areas of rough and smooth endoplasmic reticulum. There was also a marked augmentation of triglyceride-like inclusions, and later of secretory vesicles which released their content into enlarging extracellular reservoirs. This indicated a markedly high activity in the gland cells concomittant with the period of rapid synthesis of large amounts of proteins and lipoproteins destined for the growing intra-uterine larva. Mitochondrial parameters also increased, probably to generate adequate energy for synthesis, transport and secretion of nutriments for the larva. After larviposition, a marked reduction of cellular parameters was measured, and the volume density of lysosome-like structures had increased, indicating that larviposition is followed by degradation of large amounts of cellular organelles involved in biosynthetic processes. Morphometrical investigation of the uterine gland cells demonstrated a temporal correlation of the cellular dynamics with physiological events involved in producing a fully developed third-instar larva at the end of a pregnancy cycle.     

Utilization of U-14C amino acids or U-14C protein by adult Glossina morsitans during in utero development of larva

Administration of U¹⁴C protein hydrolysate in the diet of adult female Glossina morsitans at different times throughout the second reproductive cycle was followed by analysis of the distribution of radioactivity between the adult flies, their excreta, and the fully grown third instar larvae produced by these flies. A constant proportion of the total administered label was recoverable independently of the time lapse between administration and assay. Peak incorporation of labelled material occurred in the larva between the seventh and eighth day of a 9 or 10 day interlarval period, indicating that the larva feeds avidly on recently synthesized maternal uterine gland secretion at this time. Haemocoelic injection of U¹⁴C protein hydrolysate into similar adult females, between feeds, resulted in continued incorporation of labelled material by the larva to within 12 hr of parturition. Results are consistent with the hypothesis that uterine gland secretion and larval feeding continue throughout the intrauterine life of the larva.A constant and low proportion of detectable label remained in the adult fly while increased incorporation by the larva was paralleled by a reduction of detectable label in the adult excreta. This indicates direct competition between the uterine gland cells and those of the Malpighian tubules for free amino acids in the haemolymph.Administration of U¹⁴C protein in the adult diet did not result in incorporation of label by the developing larva, and the bulk was excreted as protein by the adult fly. Apparently the midgut trypsin of G. morsitans is incapable of splitting this labelled protein.Analysis of urine and haemolymph samples from flies in early pregnancy, recently fed on a diet containing U¹⁴C protein hydrolysate or U¹⁴C protein, shows that free labelled amino acids in the diet enter the adult haemolymph almost immediately after feeding, and are excreted along with dietary water during initial diuresis. The labelled protein used in these experiments was not taken up by the haemolymph and consequently did not appear in the urine.Implications are that the adult female G. morsitans possesses little storage capacity for substances in the diet which are destined to provide nutrients for the developing larva. Assuming a 48 hr digestion time, the digestive products of a blood meal ingested on day 5 or 6 of a 9 day interlarval period will provide the bulk of nutrients for larval growth. It is therefore significant that blood meals ingested at this time are larger than those ingested earlier or later in the cycle.     

Evidence for a role of progesterone in the control of uterine ornithine decarboxylase in the pregnant hamster

Changes in uterine ornithine decarboxylase (ODC) activity throughout pregnancy and the importance of progesterone in the regulation of this enzyme during the early post-implantation period have been studied in the hamster. Soon after implantation, from day 5 to day 6 of pregnancy, ODC activity rapidly increased. It reached a plateau on day 7, then abruptly fell on day 8 and remained low until the end of pregnancy. DL-alpha-difluoromethyl-ornithine (DFMO), an irreversible inhibitor of ODC, induced pregnancy termination as a consequence of the reduction of uterine ODC activity. When pregnancy arrest was induced by removing endogenous progesterone by administration of prostaglandin F2 alpha (PGF2 alpha) or by ovariectomy, the ODC rise was completely abolished, and exogenous progesterone was able to entirely counteract this effect on the enzyme activity and the termination of pregnancy. These results suggest that progesterone play a significant role in the rise of uterine ODC activity, which appears to be essential for the early post-implantation events needed for pregnancy maintenance.     

Alterations in uterine and placental prostaglandin F and E with gestational age in the rat

Experiments were designed to determine the chronological alterations in placental and uterine prostaglandin F and E (PGF and PGE) during pregnancy in the rat. Pregnant rats (sperm in the vagina = day 0) were sacrified at days 15, 18,19, 20, 21 and delivery (day 21 ) and placental and uterine tissues assayed (RIA) for PGF and PGE immediately (“ ”) or after 1 hour incubation (“ ”). Uterine content of PGF and PGE (ng PG/mg DNA) was increased significantly by day 19 and further increases were seen through delivery. Incubation of uterine tissue resulted in enhanced net production of PGF and PGE (p <.05) per mg DNA (as judged by tissue content and release into the incubation medium) by day 18 of pregnancy vs. day 15. Net production peaked around the time of delivery thus paralleling the alterations in tissue content .By contrast, no differences with gestational age were found in placental content of PGF and PGE , the concentrations throughout late gestation remaining in the range of uterine PGs at day 15. However, production of PGs per mg placental DNA increased markedly during incubation with significant enhancement detected by day 19 vs. 15, achieving levels even greater than the uterus .The and findings for the uterus are consistent with the hypothesis that increases in uterine PGs levels at the end of pregnancy may play an important role in parturition. The experiences with placental tissue suggest that the potential for PG production per placental cell may also increase in late gestation and thereby contribute to the augmented intrauterine availability of PGs at that time.     

Influence of diet on synthesis and utilisation of lipids for reproduction by the tsetse fly Glossina morsitans

Fatty acid composition of lipids from adult Glossina morsitans was unaffected by an in vitro fed diet of cow blood which induces production of under-sized offspring compared to that of flies fed on a superior diet of pig blood. The commonest fatty acids were C_16:0, C_16:1 and C_18:1 and only small differences in proportions were detected between virgin and pregnant females. Rate of lipid accumulation by males and females was the same and was unaffected by diet, but males achieved a maximum of 2.5 mg on day 9 while both virgin and fertilised females reached a maximum of 5,0 mg on day 14 of adult life. Lipid content of pregnant flies then fell to 3.0 mg on the day of larviposition and accumulation began again. A cow blood diet reduced the extent to which the lipids were utilised for larval growth and this was reflected in an altered secretory activity cycle in the female uterine gland. However, no effect on the growth of adult fat body was detectable in such flies. Mating and fertilisation, which influence reproductive events through activity of the endrocine system do not seem to affect the acquisition of lipid reserves by female Glossina. However, they clearly exert considerable influence over distribution of such reserves between fat body and uterine gland, which distribution is also affected by diet.     

Aspects of the nutrition of adult female Glossina morsitans during pregnancy

The adult female Glossina morsitans fed on goat takes, in terms of dry weight, approximately 37·6 mg of blood during a pregnancy period, and at least 84% of this dry blood comprises proteins. In this diet leucine is the most abundant, isoleucine and methionine contents are markedly small, while values for other amino acids differ and are intermediate. Shortly after pupariation the third instar larva produced by goat-fed females contains 67·3% water, 15·3% residual dry weight, 11·0% fat, and 6·4% puparium. The amount of each of the amino acids present in the larva and its puparium comprises a small proportion of that taken by the female fly during a pregnancy period. The fat content of the fully-fed larva is also very small compared with the total nutrient intake by the pregnant female. These results are discussed in terms of the nutrition of intra-uterine larva in relation to feeding by its female parent.     

The uptake of [3H]uridine into the nucleic acids of rat uterus during early pregnancy

1. Changes in content and uptake of [(3)H]uridine into the nucleic acids of rat uterus during the first 9 days of pregnancy were studied. 2. From day 6 implantation sites were separated from the rest of the uterine tissue for independent analysis. 3. Up to day 5 of pregnancy no changes were found in the total dry matter or in RNA and DNA content/unit dry matter nor in the RNA/DNA ratios. 4. From day 6, when implantation sites are visible, the water content of the implantation sites increased by 2-3%, and the RNA content/unit dry wt. and the RNA/DNA ratios increased. The DNA content/unit dry wt. did not increase in the implantation sites until day 8. 5. Uptake of [(3)H]uridine into the acid-soluble fraction of the tissues was markedly higher in implantation sites than in non-implantation sites. 6. Uptake of [(3)H]uridine into RNA was significantly increased on day 3 of pregnancy and again on day 5. 7. On days 6 and 7, the incorporation into RNA of implantation sites was significantly higher than in the remainder of the uterine tissue but decreased on days 8 and 9 to the same value as that of the normal tissue. 8. No change occurred in uptake into DNA until day 6, when there was an increase in uptake by the implantation sites. 9. It is suggested that the increase in RNA synthesis on day 3 is a preparation of the uterus for the onset of implantation on day 5, and that increased synthesis in implantation sites on days 6 and 7 is the elaboration of new RNA necessary for this early stage of pregnancy to commence.     

Activity patterns in Glossina longipennis : a field study using different sampling methods

Abstract .Studies on the daily activity of Glossina longipennis at Galana Ranch using a black odour-baited electrocuting target confirmed its crepuscular activity profile. Activity started at 05.00–05.30 hours and peaked at 06.00–06.30 hours, stopped by 09.00 hours, then started again at 17.00–17.30 hours with a peak at 18.30–19.00 hours, ceasing by 19.30 hours. Females made up 60% of the overall catch, and tended to arrive later than males. Other stationary sampling methods (trap, stationary ox) gave similar results. With the stationary methods, very few flies were caught outside the periods of peak activity (only 1.5% of the total between 09.00 and 17.00 hours); the ox was the only stationary bait to catch any flies between 10.00 and 16.00 hours. More flies were caught throughout the day at mobile baits (8.3% of the male and 2.3% of the female catch was taken between 09.00 and 17.00 hours). Mobile baits caught considerably more males than females (females were 17% of the catch). These males had on average higher fat and haematin reserves. Similar nutritional differences were not observed for females. There were fewer older females (ovarian category 3 or more) in mobile compared to stationary baits, and a lower proportion of the youngest males (wing fray category 1) at natural compared with artificial baits.     

Reproductive rates of tsetse flies in the field in Zimbabwe

Abstract. Tsetse flies Glossina morsitans morsitans Westwood and G. pallidipes Austen were marked and released within 12 h of emergence at Rekomitjie Research Station, Zambezi Valley, Zimbabwe, and on Redcliff Island, Lake Kariba. Ovarian dissections were performed on recaptured flies and on wild collected samples. At Rekomitjie >90% of female G. m. morsitans were inseminated by age 4 days and G. pallidipes by 7 days. For both species at both sites the length of the largest oocyte, for flies in ovarian category zero, increased approximately linearly for about the first 6 days and was ovulated at c. 6–8 days. The largest oocyte grew significantly more slowly in later cycles. For G. m. morsitans , but not for G. pallidipes , the rate increased with temperature; the rates were always higher than observed in the laboratory. At Rekomitjie, for both species and at a mean screen temperature of 22C, the first larva was produced at c. 18 days and subsequent larvae at 11–day intervals; the intervals decreased with temperature by c. 0.5 days/C. On Redcliff Island the intervals for both species were 2 days shorter than at Rekomitjie at any given screen temperature and were sometimes as short as 7 days. The length of the larva in utero increased exponentially during pregnancy.     

Factors affecting the landing and feeding responses of the tsetse fly Glossina pallidipes to a stationary ox

In Zimbabwe, studies were made of the landing and feeding responses of Glossina pallidipes on an ox. Of the tsetse approaching an ox, ≈ 70% fed. Increasing densities of tsetse increased the grooming responses of the ox but had no significant effect on the percentage of tsetse that engorged. The landing site of tsetse on the ox varied with density, with ≈ 50% landing on the legs at low densities (< 20 flies per ox), compared to ≈ 80% at densities > 40 flies per ox. For male G. pallidipes , the mean bloodmeal size was 37 mg. The probability of feeding was negatively correlated with fat content, declining from 91% for flies with < 1 mg fat to < 50% for flies with > 4 mg fat. Bloodmeal size was also negatively correlated with fat content; the regression equation relating bloodmeal size and fat content indicated that the mean wet weight declined from 42 mg for flies with 1 mg of fat to 31 mg for flies with 5 mg of fat. For females, the probability of feeding was not significantly affected by age as determined by ovarian category but there was a paucity of young (ovarian category 0) flies attracted to the ox. Pregnancy status had no significant effect on the probability of feeding, but samples of flies attracted to the ox showed a relative dearth of females approaching larviposition and a preponderance just after.     

Changes in mammary fat pad composition and lipolytic capacity throughout pregnancy

Changes in rat mammary fat pad during pregnancy were assessed by studying differences in the morphology and composition of the pad and in the levels of proteins involved in the accumulation and mobilization of fat stores. During pregnancy, the mammary fat pad weight had increased 1.8-fold by day 20, as compared with control rats. DNA content had increased two-fold by day 13 and remained stable until day 20. Protein content showed a two-fold increase on day 20, compared with control rats. As pregnancy advanced, both the percentage of mammary gland cells with respect to the whole mammary fat pad and the size of the adipocytes increased. The specific content of the different elements of the lipolytic pathway, viz. (α_2A-adrenergic receptor (AR), β₃-AR, cAMP-dependent protein kinase and hormone-sensitive lipase (HSL)) underwent a decrease as pregnancy progressed, although adenylate cyclase increased greatly. The lipoprotein lipase (LPL) content per gram of tissue increased with pregnancy and the HSL-to-LPL ratio reflected a continuous increase in the triglyceride storage throughout pregnancy. Thus, the mammary fat pad undergoes extensive morphological, compositional and metabolic transformation during pregnancy, attributable to the development of the mammary gland. The various elements of the lipolytic pathway and LPL undergo major changes during the development of the mammary gland focused towards the increase of fat stores and allowing the accumulation of lipid droplets in the epithelial mammary cells and an increase in adipocyte size. This investigation was supported by the Fondo de Investigaciones Sanitarias (PI021339) of the Spanish Government and by the Conselleria d'Innovació i Energia de la Comunitat Autònoma de les Illes Balears (PRDIB–2002GC4–24). E.P. was supported by a grant from the Spanish Government.     

Reproductive under-performance of tsetse flies in the laboratory, related to feeding frequency

Abstract. The rates of development of the eggs and larvae in utero and the next two developing ovarioles were measured by ovarian dissection on each day of the pregnancy cycle in tsetse, Glossina morsitans, subject to different feeding regimes. Compared with flies fed four times per pregnancy cycle, flies fed three times per cycle showed a lower pupal production rate (70%), the same (zero) adult mortality, a slightly slower growth rate of the larva and second ovariole only from day 8 onwards, but the same growth rate of the first ovariole. Flies fed only twice per pregnancy cycle produced no pupae, suffered 18% adult mortality and showed a significantly slower growth rate of the larva and second ovariole from days 6 and 7 respectively, but still the growth rate of the first ovariole was barely affected. Flies offered food three times or twice per pregnancy cycle engorged fully at every opportunity, but 16.5% of the flies offered food four times per cycle did not feed on every occasion, while 12–22% did not engorge fully on days 3, 5 or 7. In assessing the applicability of these laboratory results to the field situation the following points must be borne in mind: in the laboratory flies take smaller mean blood meals than in the field; during protein production associated with larval growth the proportion of the blood meal lost to transformation and excretory costs is less than during normal lipid metabolism; the balance between the tsetse's known fertility rate and adult and pupal mortality rates reveals that the abortion rate in the field must be extremely low. The high abortion rates usually observed in laboratory colonies, even when flies are offered food daily_l would be quite untenable in the field and indicate that laboratory conditions impose physiological stresses on the flies that are quite different from those in the field. These facts indicate that three field-sized meals may be sufficient to meet the energy demands of normal larval development in the field.