Nutritional stress affects the tsetse fly's immune gene expression.

Tsetse-transmitted trypanosomiasis poses a serious threat to human and animal health in sub-Saharan Africa. The majority of tsetse flies (Glossina spp.) in a natural population will not develop a mature infection of either Trypanosoma congolense or Trypanosoma brucei sp. because of refractoriness, a phenomenon that is affected by different factors, including the tsetse fly's immune defence. Starvation of tsetse flies significantly increases their susceptibility to the establishment of a trypanosome infection. This paper reports the effects of nutritional stress (starvation) on (a) uninduced baseline levels of gene expression of the antimicrobial peptides attacin, defensin and cecropin in the tsetse fly, and (b) levels of expression induced in response to bacterial (Escherichia coli) or trypanosomal challenge. In newly emerged, unfed tsetse flies, starvation significantly lowers baseline levels of antimicrobial peptide gene expression, especially for attacin and cecropin. In response to trypanosome challenge, only non-starved older flies showed a significant increase in antimicrobial peptide gene expression within 5 days of ingestion of a trypanosome-containing bloodmeal, especially with T. brucei bloodstream forms. These data suggest that a decreased expression of immune genes in newly hatched flies or a lack of immune responsiveness to trypanosomes in older flies, both occurring as a result of fly starvation, may be among the factors contributing to the increased susceptibility of nutritionally stressed tsetse flies to trypanosome infection.

Age prevalence of trypanosomal infections in female Glossina morsitans morsitans (Diptera: Glossinidae) on the plateau area of eastern Zambia.

Trypanosomal infections in female Glossina morsitans morsitans were investigated in an area in the Eastern Province of Zambia between 1992 and 1994. A total of 4416 flies were captured, aged using the ovarian ageing method and screened for trypanosomal infections in both the mouthparts, salivary glands and the midgut. Congolense-type infections were identified in 4.8% of the flies. Vivax-type and immature infections were identified in 1.8% and 6.8% of the flies, respectively. The prevalence of congolense-type, vivax-type and immature infections increased with age. For vivax-type infections the age-prevalence relationship could be described by a model assuming a constant per capita rate of infection. For congolense-type and midgut infections, a polynomial term was added to the model significantly improving the fit. The per capita at which flies become infected was significantly higher for immature compared to mature infections. Observations strongly suggest that tsetse acquire new midgut infections at any age and that maturation of these infections is not limited to those obtained during the first blood meal.

Effect of isometamidium chloride treatment on susceptibility of tsetse flies (Diptera: Glossinidae) to trypanosome infections.

Experiments were conducted to determine the effect of a single isometamidium chloride treatment of teneral tsetse flies, Glossina morsitans morsitans Westwood (Diptera: Glossinidae), on the subsequent susceptibility to an infection with Trypanosoma congolense or Trypanosoma brucei brucei. Flies were offered a first bloodmeal on sterile gamma-irradiated defibrinated bovine blood that contained either 10 or 100 microg ofisometamidium chloride/ml. Treated flies were subsequently infected with T. congolense IL 1180 or T. b. brucei AnTAR1 on day 3, 5, 10, or 20 posttreatment. To determine the effect of a single treatment with isometamidium chloride at 10 microg/ml on the fly's susceptibility to infection with isometamidium chloride-resistant trypanosome strains, treated flies were infected with one of two resistant isogenic T. congolense IL 1180 strains 3 d after the first feed. Results showed that a single isometamidium chloride treatment at 10 microg/ml blood sufficed to reduce significantly the fly's subsequent susceptibility to infection. Only 6.8% of the flies that were treated with isometamidium chloride developed a mature infection with T. congolense in the mouthparts compared with 34.3% of the control group. None of the flies that were administered isometamidium chloride and subsequently infected on day 3 or 6 with T. b. brucei developed a metacyclic infection in the salivary glands compared with 22.7% of the control flies. Likewise for the resistant T. congolense strains, a single treatment with isometamidium chloride significantly reduced the subsequent susceptibility to infection (6.5 and 33.5% of flies with metacyclic infections for treated and untreated flies, respectively). In practice and with respect to the release of sterile male flies to eradicate an isolated tsetse fly population, our results show that administering isometamidium chloride during the first bloodmeal (and before release) would significantly reduce the ability of these released males to transmit trypanosomes.

The transmissibility of Trypanosoma congolense seems to be associated with its level of resistance to isometamidium chloride.

In large parts of Africa the control of livestock trypanosomiasis relies on the use of trypanocidal drugs. Resistance against the available compounds is developing rapidly in the trypanosome population. The effect of the development of drug resistance on the fitness of the trypanosome is not well known. To determine the effect of the development of resistance to isometamidium chloride on the trypanosome's transmissibility, transmission experiments were conducted. Use was made of three isogenic clones of Trypanosoma congolense with different susceptibility to the drug. The infection rate in Glossina morsitans morsitans differed significantly between clones and was significantly higher in tsetse flies infected with the T. congolense clone with the highest level of drug resistance.

Ability of trypanosome-infected tsetse flies (Diptera: Glossinidae) to acquire an infection with a second trypanosome species.

The epidemiology of human and animal trypanosomiasis is determined to a large extent by the number of infected tsetse flies in a specific area. In the field, a substantial proportion of infected flies carry mixed trypanosome infections. The way in which these tsetse flies acquire a mixed infection is not fully understood. In particular, the susceptibility of tsetse flies to sequential infection with trypanosomes is not well understood. Accordingly, laboratory studies were made of the effects of age and prior infection on the probability of Glossina morsitans morsitans (Westwood) developing an infection of Trypanosoma congolense and Trypanosoma brucei brucei after feeding on infected mice. Results of these experiments clearly showed that 20-30-d-old G. m. morsitans can still pick up and develop a mature infection in the mouthparts/hypopharynx for T. congolense or in the salivary glands for T. b. brucei. However, their ability to acquire infection was significantly lower compared with teneral flies. Furthermore, 20-30-d-old flies that already carry a mature T. congolense or T. b. brucei infection remained at least as susceptible to a secondary trypanosome infection compared with noninfected flies of the same age. The immunological and epidemiological repercussions of those findings are discussed.

Comparison of the transmissibility of Trypanosoma congolense strains, isolated in a trypanosomiasis endemic area of eastern Zambia, by Glossina morsitans morsitans.

Transmission experiments were conducted to compare the transmissibility of genetically different Trypanosoma congolense (Savannah subgroup) strains isolated from cattle in a trypanosomiasis endemic area of eastern Zambia. A total of 17 strains were compared. Three strains were extremely virulent with a short pre-patent period, high parasitaemia and a short median survival time (between 5 and 9 days) in mice. The remainder of the strains belonged to the moderate (6 strains) or low (8 strains) virulence categories with median survival times between 10 and 30 days and >30 days, respectively. Batches of 40 teneral Glossina morsitans morsitans (Diptera: Glossinidae) were offered a single bloodmeal on mice infected with one of those strains. Flies were dissected to determine their infection status 21 days later. The proportion of flies with procyclic and metacyclic infections differed significantly between trypanosome strains and were significantly higher in flies infected with extremely virulent strains (P=0.033 and P=0.016 for the differences in the procyclic infection rate of strains with moderate and low virulence, respectively and P=0.005 and P=0.019 for the differences in the metacyclic infection rate of strains with moderate and low virulence, respectively). On the other hand, moderately virulent strains had, in general, higher procyclic and metacyclic infection rates compared to low virulent strains. But the differences were not significant (P>0.05). The outcome of those experiments shows clear differences in transmissibility of trypanosome strains associated with their virulence. This observation confirms the theory for the evolution and maintenance of virulence in a parasite population and may explain the persistence of virulent trypanosome strains in a susceptible host population.

The effect of starvation on the susceptibility of teneral and non-teneral tsetse flies to trypanosome infection.

Transmission of vector-borne diseases depends largely on the ability of the insect vector to become infected with the parasite. In tsetse flies, newly emerged or teneral flies are considered the most likely to develop a mature, infective trypanosome infection. This was confirmed during experimental infections where laboratory-reared Glossina morsitans morsitans Westwood (Diptera: Glossinidae) were infected with Trypanosoma congolense or T. brucei brucei. The ability of mature adult tsetse flies to become infected with trypanosomes was significantly lower than that of newly emerged flies for both parasites. However, the nutritional status of the tsetse at the time of the infective bloodmeal affected its ability to acquire either a T. congolense or T. b. brucei infection. Indeed, an extreme period of starvation (3-4 days for teneral flies, 7 days for adult flies) lowers the developmental barrier for a trypanosome infection, especially at the midgut level of the tsetse fly. Adult G. m. morsitans became at least as susceptible as newly emerged flies to infection with T. congolense. Moreover, the susceptibility of adult flies, starved for 7 days, to an infection with T. b. brucei was also significantly increased, but only at the level of maturation of an established midgut infection to a salivary gland infection. The outcome of these experimental infections clearly suggests that, under natural conditions, nutritional stress in adult tsetse flies could contribute substantially to the epidemiology of tsetse-transmitted trypanosomiasis.