Assessment of temporal changes in haem and protein levels in Ixodid ticks following blood engorgement, and their use as age-grading parameters

Nymphs of Rhipicephalus appendiculatus and Hyalomma anatolicum anatolicum were fed on rabbits and maintained in the laboratory to allow moulting and further development. At specified time intervals from 0 to 500 days after engorgement, samples of ticks were ground individually in distilled water within the wells of an agglutination plate. A 0.1 ml aliquot was removed from each and levels of haemin and protein assessed from optical density values at specific wavelengths in a spectrophotometer.

Both protein and haemin levels showed an initial rapid decrease after engorgement; values did not fall to zero in either case but showed marked fluctuation throughout the study period. These fluctuations combined with the high standard errors of the results, made assessments of the physiological age of individual ticks impossible.

Such fluctuating values, however, suggested the possibility that haem biosynthesis might be taking place, and this was tested by injecting the radioactively-labelled haem precursor [4-¹⁴C]δ-amino laevulinic acid into engorged nymphs, immediately following their detachment. Both tick species revealed an incorporation of this compound into their haematin content, although neither incorporated the non-haem precursor [1-¹⁴C]2-amino isobutyric acid. These results indicate an ability of ticks to synthesize haem in vivo, although the underlying reasons for such a mechanism remain unknown.     

Survival of unfed Rhipicephalus appendiculatus and Rhipicephalus zambeziensis adults

Two ixodid tick species, Rhipicephalus appendiculatus (Neumann) and Rhipicephalus zambeziensis (Walker et al.) (Acari: Ixodidae), both originating from the southern province of Zambia, were used to study the survival time of adults at a range of different humidities and temperatures. In general, the experiment clearly demonstrates the different survival times of the two species in relation to the climatic conditions tested: survival of R. zambeziensis was better under more extreme conditions of temperature and humidity. These findings offer an explanation for the different distribution ranges of the two tick species. Rhipicephalus appendiculatus is more confined to cooler and wetter conditions, whereas R. zambeziensis is adapted to hotter and drier areas.     

Evaluation of Food Additives and Low-toxicity Compounds for the Control of Bean Rust and Wheat Leaf Rust

The efficacy of low‐toxicity chemicals as possible alternatives to synthetic fungicides for the control of Uromyces appendiculatus and Puccinia triticina was evaluated. A preliminary selection of food additives was performed through in vitro and in vivo preliminary screenings. The ED₅₀ and minimum inhibition concentration (MIC) values showed that most of the food additives used in this study were more toxic to U. appendiculatus than to P. triticina. Acetic acid, potassium carbonate, sodium carbonate and sodium molybdate were the food additives that were more toxic to the urediniospores of P. triticina. Selected compounds and concentrations were tested on bean and wheat plants grown in pots under controlled conditions. Acetic acid, ammonium bicarbonate, potassium acetate, potassium benzoate, potassium bicarbonate, potassium carbonate, sodium acetate and sodium citrate at 0.03, 0.09, 0.03, 0.006, 0.012, 0.012, 0.03 and 0.03 m , respectively, significantly reduced the disease severity of U. appendiculatus without causing any injury to bean leaves. Ammonium bicarbonate, potassium bicarbonate, sodium bicarbonate and sodium citrate at 0.12, 0.03, 0.12 and 0.03 m , respectively, were the most effective in reducing the disease severity caused by P. triticina without causing any injury to wheat leaves.     

Gas Exchange and Emission of Chlorophyll Fluorescence during the Monocycle of Rust, Angular Leaf Spot and Anthracnose on Bean Leaves as a Function of their Trophic Characteristics

Measurements related to gas exchange and chlorophyll fluorescence emission were taken from healthy and diseased bean leaves with rust, angular leaf spot, and anthracnose during lesion development for each disease. The experiments were performed at different temperatures of plant incubation, and using two bean cultivars. The main effect of temperature of plant incubation was in disease development. There was no significant difference between cultivars in relation to disease development and in magnitude of physiological alterations when disease severity was the same for each cultivar. These diseases reduced the net photosynthetic rate and increased the dark respiration of infected leaves after the appearance of visible symptoms and the differences between healthy and diseased leaves increased with disease development. The transpiration rate and stomatal conductance were stable during the monocycle of rust, however, these two variables decreased in leaves with angular leaf spot and anthracnose beginning with symptom appearance and continuing until lesion development was complete. Carboxylation resistance was probably the main factor related to reduction of photosynthetic rate of the apparently healthy area of leaves with rust and angular leaf spot. Reduction of the intercellular concentration of CO₂, due to higher stomatal resistance, was probably the main factor for leaves with anthracnose. Chlorophyll fluorescence assessments suggested that there was no change in electron transport capacity and generation of ATP and NADPH in apparently healthy areas of diseased leaves, but decreases in chlorophyll fluorescence emission occurred on visibly lesioned areas for all diseases. Minimal fluorescence was remarkably reduced in leaves with angular leaf spot. Maximal fluorescence and optimal quantum yield of photosystem II of leaves were reduced for all three diseases. Bean rust, caused by a biotrophic pathogen, induced less damage to the regulation mechanisms of the physiological processes of the remaining green area of diseased leaves than did bean angular leaf spot or anthracnose, caused by hemibiotrophic pathogens. The magnitude of photosynthesis reduction can be related to the host–pathogen trophic relationships.     

Efficacy of certain chemical compounds on common bean rust disease

The aim of the present study was to evaluate the efficacy of some chemical compounds to management rust disease on common bean under greenhouse and field conditions. In vitro, the effect of four chemical compounds, i.e. phosphoric acid at concentrations 0.3 and 0.6%, benzoic acid, Bion® (BTH), and pyrocatechol at concentrations 4 and 8 mM from each, as well as fungicide, Tilt 25%, 0.25 and 0.5 cm³/l water on urediniospores germination of Uromyces appendiculatus was tested on water agar medium. All these compounds decreased the germination of urediniospores percentage. Under greenhouse and field experiments, application of these compounds led to decrease in disease incidence compared to the control treatments. Benzoic acid at 2015 growing seasons was the most effective in reduction of disease incidence more than the other compounds, whereas the disease incidence decreased from 59.92 to 21.43% and from 68.43 to 18.41% under greenhouse and field conditions, respectively.     

East Coast Fever of Cattle: 60Co Irradiation of Infective Particles of Theileria parva

SYNOPSIS. Infective particles (IPs) of Theileria parva , the causative organism of East Coast Fever of cattle, were harvested from the tick vector, Rhipicephalus appendiculatus , using an in vitro feeding technic. In a ranging experiment, pairs of cattle were inoculated with aliquots of suspensions of IPs irradiated at doses of 4–137.6 krad. Doses of irradiation in excess of 8 krad appeared to destroy the parasite. In the 2nd and 3rd experiments, groups of 5 cattle were inoculated with aliquots of suspensions containing low and high concentrations of IPs respectively, irradiated at doses of 4–32 krad. In the 2nd experiment, doses of irradiation in excess of 10 krad appeared to destroy the parasite. In the 3rd experiment, at least 1 animal became infected when inoculated with an aliquot of a suspension irradiated at 16 krad. In all experiments, it appeared that increasing doses of irradiation destroyed increasing numbers of IPs. There was no conclusive evidence that IPs which survived irradiation were attenuated, and it appears that vaccination of cattle against ECF is unlikely to be achieved by inoculation of irradiated IPs using the methods described.     

The incorporation of radio-labelled nucleic acid precursors by Theileria parva in bovine blood and salivary glands of Rhipicephalus appendiculatus ticks

Irvin A.D., Boarer C.D.H., Kurtti T.J. and Ocama J.G.R. 1981. The incorporation of radio-labelled nucleic acid precursors by Theileria parva in bovine blood and salivary glands of Rhipicephalus appendiculatus ticks. International Journal for Parasitology11:451–456. The uptake of radio-labelled nucleic acid precursors by blood and tick salivary gland forms of Theileria pana was studied. Piroplasms took up tritiated purines, particularly hypoxanthine, but not pyrimidines. Similar uptake was recorded by T. parva, both in tick saliva and in salivary glands maintained in vitro. Intermediate parasite stages were those most readily labelled in glands; this reflected active nucleic acid synthesis associated with rapid parasite division. Radio-labelling of T. parva in tick salivary glands could be of value in procedures used for concentrating and purifying theilerial sporozoites.     

Effects of defoliation on the saplings of a gap-colonizing neotropical tree

This paper describes a defoliation experiment on saplings of the gap-colonizing pioneer tree Heliocarpus appendiculatus in the tropical rain forest of Los Tuxtlas (Southeast Mexico). Four levels of defoliation (0, 25, 50, and 75% of the leaf area removed) were applied to naturally established plants in a medium-sized forest gap. Records were made of growth (height, diameter, and leaf production) and of whole-plant and leaf survival. Statistical comparisons for all variables showed that plants that had not been defoliated grew better than defoliated ones, but there were no statistically significant differences between the defoliation levels. The risk of death was significantly lower for control plants than for defoliated plants at any level of damage. Leaf survivorship was highly irregular. The most consistent pattern was that the leaves of intact plants always showed higher survivorship, while the most heavily defoliated ones always had the poorest survival. The survivorship pattern of leaves at intermediate defoliation levels was irregular. The results illustrate the lack of a monotonic response to a wide range of defoliation levels, and suggest the potential effect of herbivores as reducers of vegetative growth and survival in pioneer tropical species, and as limiting agents of plant establishment in regenerating forest gaps.     

Identification of a complex peptidergic neuroendocrine network in the hard tick, <Emphasis Type="Italic">Rhipicephalus appendiculatus</Emphasis>

Neuropeptides are crucial regulators of development and various physiological functions but little is known about their identity, expression and function in vectors of pathogens causing serious diseases, such as ticks. Therefore, we have used antibodies against multiple insect and crustacean neuropeptides to reveal the presence of these bioactive molecules in peptidergic neurons and cells of the ixodid tick Rhipicephalus appendiculatus. These antibodies have detected 15 different immunoreactive compounds expressed in specific central and peripheral neurons associated with the synganglion. Most central neurons arborize in distinct areas of the neuropile or the putative neurohaemal periganglionic sheath of the synganglion. Several large identified neurons in the synganglion project multiple processes through peripheral nerves to form elaborate axonal arborizations on the surface of salivary glands or to terminate in the lateral segmental organs (LSO). Additional neuropeptide immunoreactivity has been observed in intrinsic secretory cells of the LSO. We have also identified two novel clusters of peripheral neurons embedded in the cheliceral and paraspiracular nerves. These neurons project branching axons into the synganglion and into the periphery. Our study has thus revealed a complex network of central and peripheral peptidergic neurons, putative neurohaemal and neuromodulatory structures and endocrine cells in the tick comparable with those found in insect and crustacean neuroendocrine systems. Strong specific staining with a large variety of antibodies also indicates that the tick nervous system and adjacent secretory organs are rich sources of diverse neuropeptides related to those identified in insects, crustaceans or even vertebrates. This work was supported by Slovak grant agencies: Agentúra na podporu vyskumu a vyvoja (APVV-51-039105) and Vedecká grantová agentúra (VEGA 2-6090-26 and 2/6155/26).     

Molecular individuality and adaptation of the tick Rhipicephalus appendiculatus in changed feeding environments

The tick Rhipicephalus appendiculatus Neumann (Acari: Ixodidae) naturally infests many host species. However, the mechanisms that enable it to feed on such a wide range of hosts are unclear. One possibility is that a tick population maintains molecular (genotypic and/or phenotypic) diversity among individuals such that individuals vary in their competency in taking bloodmeals under different feeding conditions. As a first step in testing this hypothesis, we showed that the polymorphism of salivary gland proteins, previously demonstrated in unfed ticks, was maintained during feeding on guinea-pigs. We then compared feeding performance under standard laboratory rearing conditions: one instar (adults or nymphs) feeding on guinea-pigs, with three changed conditions: (1) two instars (adults and nymphs) feeding together on guinea-pigs; (2) one instar (adults or nymphs) feeding on hamsters; and (3) two instars (adults and nymphs) feeding together on hamsters. The mean engorged weight of adult females was significantly reduced under all changed conditions, indicating that most of the adult individuals were significantly challenged by the changed conditions. However, some individuals achieved successful engorgement, indicating competence to the changed condition, and demonstrating variation in adaptive ability among individuals. Engorged females produced egg masses positively correlated to the engorged weights. More interestingly, the correlation coefficient (R) increased when feeding condition was changed. This may lead to more efficient selection for population adaptation under the changed conditions. As the feeding success of ixodid ticks depends on the efficiency of the cocktail of immunomodulatory saliva, the relevance of the polymorphism of salivary gland proteins and host adaptation is discussed.