X-ray investigation of gas bubble formation, and water loss in tsetse fly pupae

ABSTRACT. Using a microfocal X-ray apparatus, a gas bubble was detected within the puparium of Glossina morsitans. The bubble appeared between 6 and 15 h after pupariation and was associated with one of the longitudinal tracheal trunks of the third instar larva. The bubble grew and achieved maximum size approximately 96 h after pupariation. It then disappeared at the time of eversion of the pupal appendages. There was a close correlation between bubble size and the weight of water lost since the time of pupariation. At the time of eversion of the pupal appendages the gas bubble apparently passed out through the longitudinal tracheal trunk and posterior spiracle to occupy the space between larval (puparial) and pupal cuticle. It is suggested that the bubble plays a vital role in the separation of these cuticular layers and that to this end water loss from the puparium is essential.     

Mortality of olive fruit fly pupae in California

Efforts to control the olive fruit fly, Bactrocera oleae (Diptera: Tephritidae), in California have focused on insecticidal baits and biological control by parasitoids, which primarily target the adult and larval stages, respectively. The pupal stage, which occurs in the soil, has largely been overlooked. This study investigated mortality factors for olive fruit fly pupae in California olive orchards, using a combination of exclusion experiments and observation and trapping of potential predators. Results show predation and climatic factors contribute to pupal mortality. Ants (Formicidae) were the most numerous predators observed. Soil-borne pathogens caused no mortality in this study. Potential applications of these results in the development of a sustainable management program are discussed.     

Glycogen in the proventriculus of the tsetse fly

Glycogen was detected in the proventriculus of the tsetse fly, Glossina morsitans morsitans, by ultrastructural, histochemical, and biochemical methods. This organ contained ten times or more glycogen on a dry weight basis than was found in the thoracic muscle. Proventriculi of male tsetse contained less glycogen than those of females belonging to the same age group and in teneral flies the amount of glycogen was about 50 per cent lower than in mature, fed flies of the same sex. Although the thoracic muscle of tsetse flies was considerably lower in glycogen than that of blowflies the amounts in the proventriculus of mature females of the two insect species were almost equal. It is suggested that this carbohydrate store may supply the energy required for secretory processes.     

Differentiating tobacco budworm and corn earworm using near-infrared spectroscopy

Near-infrared spectroscopy (NIRS) was used to develop a simple and quick technique to differentiate two economically important species, the tobacco budworm, Heliothis cirescens (F.), and corn earworm, Helicoverpa zea (Boddie), which are major pests of cotton, Gossypium hirsutum L., in the southern United States. In practice, it is difficult to distinguish the two species during their immature stages using morphological characteristics unless expensive microscopy equipment or trained technicians are available. The current studies demonstrated that the two species could be quickly and readily differentiated during early developmental stages, including egg and young larval (younger than third instar) stages, by using NIRS technology with up to 95% accuracy. NIRS technology could significantly improve pest diagnosis in cotton pest management.     

Diuresis in the tsetse fly Glossina austeni.

After taking a blood meal, the tsetse fly Glossina austeni excretes the excess water and salts of the meal in approximately 30 min. During this period a volume of fluid equivalent to 80% of the unfed weight of the fly passes through the haemolymph, whose composition nevertheless remains almost constant. The fluid excreted has a higher sodium and lower potassium concentration than the haemolymph, indicating that sodium may be the prime mover in urine formation in Glossina.     

Octopamine distribution in the tsetse fly Glossina morsitans

Tissues of Glossina morsitans were assayed for octopamine using an enzymatic technique. Octopamine was detected at the highest concentration in the brain (7.06-7.99 ng mg-1 tissue protein) and thoracic ganglion (10.9-13.89 ng mg-1 tissue protein). Octopamine was present in haemolymph at a concentration of 1.0-1.27 X 10(-7) M. This was not found to vary when insects were flown or mechanically stressed. Nervous tissue, flight muscle and haemolymph showed a significant ability to metabolize octopamine. The greatest enzyme activity was present in the haemolymph.     

Micro-organisms in the midgut of tsetse fly larvae.

Two types of micro-organisms were found in the midgut of Glossina morsitans larvae, a large Gram-negative bacterial rod and a small Gram-negative rickettsia-like micro-organism, although the occurrence of the rickettsial type is restricted. The location of these micro-organisms in a small area of the proventriculus of all three larval instars is discussed. The large micro-organisms resemble milk-gland bacteria, and further evidence is presented in support of a milk transmission hypothesis for these micro-organisms.     

Microalgal fatty acid composition: rapid assessment using near-infrared spectroscopy

The feasibility of assessing microalgal fatty acid composition using near-infrared spectroscopy (NIRS) is described. The chlamydomonad microalga, Rhopalosolen saccatus (previously known as Characium saccatum), was isolated from the Fitzroy River, Central Queensland, Australia. R. saccatus was grown in batch culture with varying phosphorus nutrition and assessed for dry matter, total lipid and fatty acid composition using gas chromatography (GC). Transmission spectra (1100–2500 nm) were acquired of liquid culture, and reflectance spectra were acquired of wet and dry filtrates of cultures and of methyl esters. Partial least square (PLS) regression models were built on biomass, total lipid and a number of fatty acids. All sample presentation models supported PLS regression model with a cross validation correlation coefficient (R _cv) >0.87 for biomass and R _cv >0.68 for total lipid; however, the use of dry filtrates of culture is recommended as the sample presentation mode of choice. Models for fatty acids based on culture transmission spectra, reflectance spectra of wet and dry culture filtrates, or reflectance spectra of methyl esters in solvent were not acceptable. Dry extracts of methyl esters supported adequate models for fatty acids from C8:0 to C22:0, with the exception of capric and behenic acids, with an R _cv of 0.89–0.94; however, in practice, samples processed to this stage can be easily analyzed by GC. Near-infrared spectroscopy can be a potential choice for rapid estimation of biomass (dry matter) and lipid content and composition in microalgae, with further work required to demonstrate oping robustness of the calibration model in prediction of unknown samples.     

At-line monitoring of a submerged filamentous bacterial cultivation using near-infrared spectroscopy

The use of near infra-red spectroscopy (NIRS) to monitor a submerged filamentous bacterial bioprocess was investigated. An industrial strain of the filamentous bacterium Streptomyces fradiae was cultured in a 12 litre stirred tank reactor (STR) using a complex medium. This mycelial 4 phase (oil, water, gas and solid) system produced highly complex and variable matrices, therefore monitoring such a complex fluid with NIRS represented a considerable challenge. Nevertheless, successful models for four key analytes (methyl oleate, glucose, glutamate and ammonium) were built at-line (rapid off-line) using NIRS. In the present study, the methods used to formulate, select and validate the models for the key analytes are discussed, with particular emphasis on how the model performance can be critically evaluated. Since previous reports on NIRS in monitoring bioprocesses have either involved simpler matrices, or, in filamentous systems, have not discussed how NIRS models can be critically assessed, the emphasis in the present study on providing an insight into the modelling process in such a complex matrix, may be particularly important to the applicability of NIRS to such industrial bioprocesses.     

Genetic analysis by DNA fingerprinting in tsetse fly genomes

Genomic DNA from tsetse flies (Diptera : Glossinidae: Glossina Wiedemann) was analyzed by hybridization using the whole M13 phage as a probe to reveal DNA fingerprinting (DNAfp) profiles. Intrapopulation variablity, measured by comparison of DNAfp profiles of tsetse flies from large colony of G. brevipalpis, showed a high degree of polymorphism similar to that found in other animal species. Different lines of G. m. morsitans, G. m. centralis, G. m. submorsitans, G. p. palpalis and G. p. gambiensis established from small colonies displayed less genetic variability than the G. brevipalpis population. The analysis of pedigree relationships within an inbred line of G. m. centralis conformed to a Mendelian inheritance pattern. In the pedigree presented no mutations were observed, one fragment was linked to the X chromosome, and three fragment sets were linked, but most fragments showed independent segregation. M13 revealed no characteristics DNAfp profile differences between the subgenus Glossina and the subgenus Nemorhina, but a conserved distribution pattern was found in the laboratory colonies within each subspecies. M13 also revealed line specific DNA fragments that may be useful as genetic markers to expand the present linkage map of G. m. morsitans.     

Population vulnerability and disability in Kenya's tsetse fly habitats

Background

Human African Trypanosomiasis (HAT), also referred to as sleeping sickness, and African Animal Trypanosomaisis (AAT), known as nagana, are highly prevalent parasitic vector-borne diseases in sub-Saharan Africa. Humans acquire trypanosomiasis following the bite of a tsetse fly infected with the protozoa Trypanosoma brucei (T.b.) spp. –i.e., T.b. gambiense in West and Central Africa and T.b. rhodesiense in East and Southern Africa. Over the last decade HAT diagnostic capacity to estimate HAT prevalence has improved in active case-finding areas but enhanced passive surveillance programs are still lacking in much of rural sub-Saharan Africa.

Methodology/Principal Findings

This retrospective-cross-sectional study examined the use of national census data (1999) to estimate population vulnerability and disability in Kenya''s 7 tsetse belts to assess the potential of HAT-acquired infection in those areas. A multilevel study design estimated the likelihood of disability in individuals, nested within households, nested within tsetse fly habitats of varying levels of poverty. Residents and recent migrants of working age were studied. Tsetse fly''s impact on disability was conceptualised via two exposure pathways: directly from the bite of a pathogenic tsetse fly resulting in HAT infection or indirectly, as the potential for AAT takes land out of agricultural production and diseased livestock leads to livestock morbidity and mortality, contributing to nutritional deficiencies and poverty. Tsetse belts that were significantly associated with increased disability prevalence were identified and the direct and indirect exposure pathways were evaluated.

Conclusions/Significance

Incorporating reports on disability from the national census is a promising surveillance tool that may enhance future HAT surveillance programs in sub-Saharan Africa. The combined burdens of HAT and AAT and the opportunity costs of agricultural production in AAT areas are likely contributors to disability within tsetse-infested areas. Future research will assess changes in the spatial relationships between high tsetse infestation and human disability following the release of the Kenya 2009 census at the local level.     

Early pregnancy diagnosis in sheep using near-infrared spectroscopy on blood plasma

The objective of this study was to evaluate the ability of near-infrared reflectance spectroscopy (NIRS) to discriminate between pregnant and nonpregnant ewes in early stages of pregnancy after artificial insemination (AI) from blood plasma. Samples were collected using jugular puncture at 18 and 25 days after AI from 188 Rasa Aragonesa and Ansotana ewes. Plasma samples were analyzed for pregnancy-associated glycoprotein (PAG) and progesterone (P4) using ELISA commercial kits. The spectra of plasma samples were recorded in the visible and near-infrared ranges. The performance of these tests were compared, using as criterion standard the pregnancy status determined using transabdominal ultrasonography at 45 days after AI. Pregnancy rate was 47.9% (90/188). At Day 18, sensitivity was similar in NIRS and P4 tests (98.9% vs. 100%; not significant) and greater than PAG (32.2%; both P < 0.001). Specificity was similar in NIRS and PAG tests (both 100%) and greater than that of P4 (84.7%; P < 0.001). At Day 25, sensitivity and specificity of NIRS and PAG were both 100%. It can be concluded that NIRS was an accurate method of diagnosis of pregnancy at Days 18 and 25 after AI in ewes.