The nervous systems of Tylodelphys metacercariae (Digenea: Diplostomidae) from the catfish Clarias gariepinus (Clariidae) in freshwater habitats of Tanzania.

The nervous systems of three Tylodelphys metacercariae (T. mashonense, Tylodelphys spp. 1 and 2) co-occurring in the cranial cavity of the catfish, Clarias gariepinus, were examined by the activity of acetylthiocholine iodide (AcThI), with the aim of better understanding the arrangement of sensillae on the body surface and the nerve trunks and commissures, for taxonomic purposes. Enzyme cytochemistry demonstrated a comparable orthogonal arrangement in the three metacercariae: the central nervous system (CNS) consisting of a pair of cerebral ganglia, from which anterior and posterior neuronal pathways arise and inter-link by cross-connectives and commissures. However, the number of transverse nerves was significantly different in the three diplostomid metacercariae: Tylodelphys sp. 1 (30), Tylodelphys sp. 2 (21) and T. mashonense (15). The observed difference in the nervous system of the three metacercariae clearly separates them into three species. These findings suggest that consistent differences in the transverse nerves of digenean metacercariae could enable the differentiation of metacercariae to the species level in the absence of molecular techniques. This, however, might require further testing on a larger number of species of digenean metacercariae.

Completion of the life cycle of Tylodelphys mashonense (Sudarikov, 1971) (Digenea: Diplostomidae) with DNA barcodes and rDNA sequences.

The life cycle of Tylodelphys mashonense (Digenea: Diplostomidae), whose metacercariae occur in the cranial cavity of the widely cultivated catfish Clarias gariepinus, was resolved by the application of molecular markers. Both COI barcodes and ITS sequences obtained from diplostomid-like cercariae infecting Bulinus spp. from Mindu Dam, Morogoro, matched those acquired from metacercariae from the catfish C. gariepinus, and those from adult T. mashonense from the grey heron Ardea cinerea and the white egret Egretta alba. The success in linking the life cycle stages of T. mashonense using molecular tools highlights the usefulness of this approach in resolving the complex life cycles of digeneans in the absence of experimental establishment.

Plasmodium falciparum msp2 Genotypes and Multiplicity of Infections among Children under Five Years with Uncomplicated Malaria in Kibaha, Tanzania.

Genetic diversity of Plasmodium falciparum may pose challenges in malaria treatment and prevention through chemotherapy and vaccination. We assessed Plasmodium falciparum genetic diversity and multiplicity of infection (MOI) of P. falciparum infections and sort relationship of parasitaemia with P. falciparum msp2 genotypes as well as with the number of infecting clones. The study was carried out in Kibaha, Tanzania. Ninety-nine children under five years with uncomplicated malaria were recruited. Genetic diversity was analyzed by genotyping the msp2 gene using PCR-Restriction Fragment Length Polymorphism. Thirty-two different msp2 alleles were obtained. The msp2 3D7 allelic frequency was higher (48.1%) and more prevalent than FC27 (27.3%) (p < 0.05). Twenty-four percent of the infections were mixed alleles. The individuals with FC27 had high parasitemia compared to those with 3D7 alleles (p = 0.038). The mean MOI was low (1.4 clones, 95% CI 1.2-1.5). The P. falciparum population among children at Kibaha is composed of distinct P. falciparum clones, and parasites having 3D7 are more frequent than those with FC27 alleles. Individuals with parasite having FC27 alleles have high parasite densities suggesting that parasites with FC27 alleles may associate with severity of disease in Kibaha. Low MOI at Kibaha suggests low malaria transmission rate.

Diversity, spatial and temporal abundance of Anopheles gambiae complex in the Rufiji River basin, south-eastern Tanzania.

The Anopheles gambiae complex contains the most efficient malaria vectors in the world. Identification of the species and the concomitant distribution are vital for effective malaria control. The objective of the study was to establish the diversity, spatial and seasonal abundance of malaria vectors in the Rufiji River Basin in Tanzania and the implications on malaria vector control. Mosquitoes were collected using CDC light-traps. Anopheles mosquitoes were identified by conventional keys. Anopheles gambiae complex were speciated using standard PCR method. Of the 562 specimens analysed by PCR 69% produced fragments equivalent to An. gambiae s.s. (390 bp), 23% equivalent to An. arabiensis (315 bp), and 7% as An. merus (464 bp). An. gambiae s.s. and An. merus were more abundant on the plateau than on the flood plain (Fisher's exact test, P < 0001), whereas An. arabiensis was equally abundant between the two sites (Fisher's exact test, P=1656). The density of the three sibling species of An. gambiae complex also varied with the seasons. An. gambiae s.s. and An. arabiensis were most predominant species during the start of the rainy season, but as season progresses, An. gambiae s.s. predominated. An. merus was only recorded during the short rainy season. In conclusion, both An. gambiae s.s., An. arabiensis and An. merus are malaria vectors in the Rufiji River basin and that An. merus is recorded for the first time in the south-eastern coast of Tanzania. These findings are important in the planning and implementation of malaria vector control activities in the Rufiji River basin, south-eastern Tanzania.

Variation in the morphometrics of diplostomid metacercariae (Digenea: Trematoda) infecting the catfish, Clarias gariepinus in Tanzania.

Diplostomum species are economically important worldwide due to their metacercariae which parasitize the eyes of fish, in both natural and aquaculture systems. However, their striking morphological similarity, especially at the metacercarial stage, makes species separation difficult. Three closely related diplostomid metacercariae, namely Diplostomum mashonense (type 3), Tylodelphys sp. 1 and 2 (type 1 and 2, respectively), coexist in the cranial cavity of the catfish, Clarias gariepinus from Mindu dam, Lake Victoria, Msimbazi, Ruvu and Kilombero rivers. The morphometrics of these three species were analysed by discriminant function analysis to investigate the degree of variation among the populations from the five localities sampled. The first canonical functions for all visual examination plots accounted for over 50% of the between-group variability. The observed differences were mainly from measurements associated with length, indicating that these measurements are important in the description of population characteristics. Visual examination of the samples along the canonical functions showed a clear between-population differentiation. The overall random assignment of individuals into their original groups was high (97%). These extensive morphometric variations introduce doubt about the reliability of measurements in the determination of species in these trematodes, as they may lead to misidentifications.

Nature beats nurture: a case study of the physiological fitness of free-living and laboratory-reared male Anopheles gambiae s.l.

Laboratory experimentation forms the basis for most of our knowledge of the biology of many organisms, in particular insects. However, the accuracy with which laboratory-derived estimates of insect life history and behaviour can predict their fitness and population dynamics in the wild is rarely validated. Such comparison is especially important in cases where laboratory-derived information is used to formulate and implement strategies for the genetic control of insects in nature. We have conducted a comparative study of the reproductive potential and life history of male Anopheles gambiae Gilies sensu lato mosquitoes from both standardized laboratory conditions and from natural field settings. We measured three indirect indicators of male mosquito fitness: energetic reserves, body size and survival, in a bid to determine whether the demographics and energetic limitations of wild males can be correctly predicted from their laboratory counterparts. Crucially, the body size and lipid reserves of wild males were substantially greater than those reared under standard laboratory conditions. We caution that the energetic limitations of insects as identified in the laboratory may underestimate their resilience in the wild, and discuss the implications of this phenomenon with respect to vector-borne disease control programmes based on genetic control of mosquitoes.

Hosts of Glossina fuscipes fuscipes and G. pallidipes in areas of western Kenya with endemic sleeping sickness, as determined using an egg-yolk (IgY) ELISA.

Bloodmeal sources of Glossina fuscipes fuscipes and G. pallidipes, from the western Kenyan foci of human African trypanosomiasis (HAT) on Mageta Island and in Busia district, were identified using an ELISA based on chicken egg-yolk (IgY) antibodies. After absorption with cross-reacting antigens, the antibodies, which were produced against representatives of eight families of vertebrate host, were capable of differentiating serum from the different families. With the ELISA, it was possible to identify the family of host for 100% of laboratory-fed flies tested up to 48 h post-bloodmeal but only for 12% of such flies tested 96 h post-feed. Subsequently, attempts were made to identify the family of host that was the source of the (most recent) bloodmeal for each of 223 wild-caught flies, and these attempts were successful for 142 (63.7%) of the samples. Among the flies with identified bloodmeals, most (81.9%) of the G. f. fuscipes caught on Mageta Island had last fed on reptiles whereas most of the G. f. fuscipes (70.4%) and G. pallidipes (57.1%) caught in Busia had last fed on bovids. Bloodmeals of human origin accounted for <2% of the bloodmeals identified, perhaps indicating that, in the presence of alternative hosts, humans are not attractive hosts for tsetse in the study areas. This finding may account for the low reported incidence of HAT, despite the presence of circulating human-infective trypanosomes. In Busia at least, the use of animals, especially cattle, covered in insecticide would probably be an effective method of controlling the tsetse vectors of the trypanosomes that cause human and 'animal' trypanosomiases.

Dhfr and dhps mutations in Plasmodium falciparum isolates in Mlandizi; Kibaha; Tanzania: association with clinical outcome

Sulfadoxine-pyrimethamine (SP); the current first line antimalarial drug in Tanzania; is compromised by evolution and spread of mutations in the parasite's dhfr and dhps genes. In the present study we established the baseline frequencies of Plasmodium falciparum dihydrofolate reductase (pfdhfr) and dihydropteroate synthase (pfdhps) mutant genotypes and their potential for predicting the in vivo efficacy of SP in Mlandizi; Tanzania. The efficacy of SP treatment was by following 116 children with uncomplicated falciparum malaria for 14 days after treatment. Infected blood samples were collected on filter paper at days 0; 3; 7 and 14. Parasite genomic DNA was extracted and point mutations at positions 51; 59; 108 and 164 of the dhfr gene and at 581; 540 and 437 of the dhps gene were analysed by nested Polymerase Chain Reaction/ Restriction Fragment Length Polymorphism. Out of 116 children enrolled; 98 (86) of eligible children demonstrated an adequate clinical response by day 14. There were 7.3early and 6.7late therapeutic failures. At day 0; only 8.0(4/50) the parasites showed no mutation at the dhfr locus; for dhps this was 73. Triple mutant dhfr alleles (Ile 51; Arg 59; Asn 108) occurred in 47; double mutant dhps (Gly 437; Glu 540) alleles in 7.9. No mutation was detected at codon 164 of the dhfr gene. The presence of triple dhfr mutant alleles was related to clinical failure; but did not show significant association (Fisher exact test; P=0.166; OR 2.15 0.77OR6.20). The higher rates of mutation on the dhfr do not spell a bright future for SP treatment in Tanzania. It is rational to think of an alternative first line antimalarial drug; while retaining SP for malaria intermittent treatment in pregnancy

Evaluation of the efficacy of the crude extract of Aloe secundiflora in chickens experimentally infected with Newcastle disease virus.

Two replicate experiments were carried out to verify the efficacy of Aloe species (Aloaceae) as used for the control of Newcastle disease (ND) in rural poultry in free-range systems among several communities in Tanzania. Four months old local chickens free of Newcastle disease antibodies were used. Following inoculation with ND virus, body weights, clinical signs, antibody levels and mortality were monitored. Results showed that there was reduced mortality rate and the severity of clinical signs during the acute phase of the infection in Aloe treated chickens compared with the non-treated ones. However, there was no significant effect of the Aloe on the antibody levels that were attributed to the recovery of the surviving chickens. The findings of this study suggest that Aloe secundiflora could be a potential candidate on the management of Newcastle disease in chickens. Further studies are in progress to identify the active ingredients of A. secundiflora against Newcastle disease virus.

Efficacy of crude extract of Aloe secundiflora against Salmonella gallinarum in experimentally infected free-range chickens in Tanzania.

The ethnoveterinary use of Aloe species extract in free-range local chickens against fowl typhoid was investigated. Five-months-old local chickens, free of antibodies against fowl typhoid were used. The chickens were randomly assorted into five groups including pretreated and infected (G1, n=21), infected and untreated (G2, n=21), infected and treated (G3, n=21), untreated and uninfected (G4, n=10) and treated uninfected (G5, n=10). Groups 1, 2 and 3 were inoculated with 5.0 x 10(8) c.f.u/ml of Salmonella gallinarum, following which the chickens were monitored for 15 days. There was a delay on the occurrence of the clinical signs and reduced severity of the disease in the Aloe treated chickens (G1 and G3). The mortality rates were 23.8% (5/21) in G1 (pretreated and infected), 42.8% (9/21) in G2 (infected and untreated) and 14.2% (3/21) in G3 (infected and treated). Antibody levels were raised among the infected and untreated group (G2) while they remained relatively low in the Aloe treated groups (G1 and G3). However, there was a sharp increase in the levels of interleukin 6 (IL-6) in the Aloe treated groups (G1 and G3) as compared to the untreated group (G2) until day 9 post-infection. The results of this work indicate that the extract of Aloe secundiflora may be used in the control of fowl typhoid in chickens. Studies to determine the active ingredients of the plant extract are in progress.