In silico SNP prediction of selected protein orthologues in insect models for Alzheimer's, Parkinson's, and Huntington's diseases.

Alzheimer's, Parkinson's, and Huntington's are the most common neurodegenerative diseases that are incurable and affect the elderly population. Discovery of effective treatments for these diseases is often difficult, expensive, and serendipitous. Previous comparative studies on different model organisms have revealed that most animals share similar cellular and molecular characteristics. The meta-SNP tool includes four different integrated tools (SIFT, PANTHER, SNAP, and PhD-SNP) was used to identify non synonymous single nucleotide polymorphism (nsSNPs). Prediction of nsSNPs was conducted on three representative proteins for Alzheimer's, Parkinson's, and Huntington's diseases; APPl in Drosophila melanogaster, LRRK1 in Aedes aegypti, and VCPl in Tribolium castaneum. With the possibility of using insect models to investigate neurodegenerative diseases. We conclude from the protein comparative analysis between different insect models and nsSNP analyses that D. melanogaster is the best model for Alzheimer's representing five nsSNPs of the 21 suggested mutations in the APPl protein. Aedes aegypti is the best model for Parkinson's representing three nsSNPs in the LRRK1 protein. Tribolium castaneum is the best model for Huntington's disease representing 13 SNPs of 37 suggested mutations in the VCPl protein. This study aimed to improve human neural health by identifying the best insect to model Alzheimer's, Parkinson's, and Huntington's.

Impact of anti-sandfly saliva antibodies on biological aspects of Phlebotomus papatasi (Diptera: Psychodidae), vector of cutaneous leishmaniasis.

Sandflies are the main vectors of Leishmania parasites in tropical and subtropical areas. The immunization of vertebrate hosts with vector components through repeated bites may offer an alternative method for sandfly control. Aliquots of female Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) were weekly blood fed on 12 individual hamsters throughout 18 successive weeks. Significant biological and biochemical changes resulting from antibodies developed by immunized host sera against repeated biting were observed in sandfly females. Blood feeding and fertility rates of females significantly gradually declined to the end of the study period. No appreciable difference was observed in mortality rates among flies repeatedly fed on individual hamsters throughout weeks 9 and 18, compared to flies fed on naïve hamsters. Total salivary gland proteins of female sandflies were compared to proteins in sera of sensitized hamsters. SDS-page revealed bands common to both flies and hosts, indicating the development of anti-saliva antibodies in hamster sera. The importance of anti-sandfly saliva antibodies as a potential tool for vector control leading to the interruption of leishmaniasis is discussed.

Experimental effect of feeding on Ricinus communis and Bougainvillea glabra on the development of the sand fly Phlebotomus papatasi (Diptera: Psychodidae) from Egypt.

Plants are promising sources of agents useful for the control of vectors of human diseases including leishmaniasis. The effect of Ricinus communis (Euphorbiaceae) and Bougainvillea glabra (Nyctaginaceae), on transmission of leishmaniasis was investigated using them as diets for Phlebotomus papatasi to monitor their effect on life-history traits. P. papatasi were allowed to feed separately on both plants then offered a blood-meal. Fed-females were observed daily for egg-laying and subsequent developmental stages. P. papatasi was able to feed on B. glabra (29.41% females and 46.30% males) and R. communis (5.80% females and 10.43% males). 34.28% of females died within 24-48 hours post-feeding on R. communis, whereas, it was 16.5% in females fed on B. glabra. Overall fecundity of surviving females was reduced compared to controls, reared on standard laboratory diet; however there was no effect on the sex ratio of progeny. Female P. papatasi in the control group had significantly longer life span compared to plant-fed group. Feeding on these plants not only decreased sand fly survival rates but incurred negative effects on fecundity. Findings indicate that planting high densities of R. communis and B. glabra in sand flies-endemic areas will reduce population sizes and reduce the risk of Leishmania major infections.

Natural and experimental evidence of viscerotropic infection caused by Leishmania tropica from North Sinai, Egypt.

Cutaneous leishmaniasis (CL) is a neglected clinical form that is quite prevalent in Eastern North parts of the country in Sinai Peninsula. Leishmania tropica was identified by previous reports as the causative agent responsible for viscerotropic infections in-patients and experimental animals. Here, we reported the viscerotropic infections from naturally infected rodent Gerbillus pyramidum floweri collected from North-Sinai. Footpad and tail lesions, spleenomegaly, and malformed dark-colored spleen were the characteristic CL symptoms. The spleen of the rodent found positive to amastigote impression smear. ITS-1 DNA was sequenced and revealed 100% identity of the strain in the current study to the other L. tropica sequences identified from the patients with the suspected CL and inhabited the same study area. The current findings confirmed the susceptibility of gerbil to L. tropica, and raise the concerns for the role of rodents as accidental host suffering the infections. The susceptibility of wild and experimental rodents to the same L. tropica strain was also investigated; BALB/c and G. pyramidum were more susceptible to L. tropica (24.33 ± 4.37 and 25 ± 4.58 days post-infection, respectively). Similar viscerotropic pathologies were reported in experimental infection of only golden hamster (≈ 120 days post-infection), and G. p. floweri (≈ 160 days post-infection).

Morphometrics and protein profiles of the salivary glands of Phlebotomus papatasi and Phlebotomus langeroni sand flies.

Sand fly salivary fluid contains numerous proteins that modulate host immune responses to infection and facilitate blood-feeding and establishing Leishmania infection. Salivary proteins are differentially expressed in adaptation to the host, the meal type and ecological factors. We report on the morphometrics and protein composition of salivary glands of colonised Phlebotomus papatasi and P. langeroni sand flies from Egypt. Female glands were dissected at day 1 (D1, unfed), day 2 (D2, sugar-fed), day 3 (D3, blood-fed) and day 7 (D7, blood-digested). The salivary glands are composed of two lobes: heterogeneous in P. papatasi and homogeneous in P. langeroni. Lobe sizes varied considerably with fly age and feeding state; D3 flies had the largest lobe sizes and protein content. The P. papatasi flies had larger lobes and higher protein content than the P. langeroni flies. The P. papatasi D1 flies had 15 protein bands that decreased in the D2, D3 and D7 flies to 10 bands in the Sinai flies and 9 bands in the Alexandria flies. All P. langeroni flies had 12 protein bands but with different intensities. The results reveal inter-specific variation between P. papatasi and P. langeroni, while no intra-specific variation between P. papatasi strains. These results increase our understanding of salivary gland protein composition and blood-feeding behaviour in Old World sand flies with implications for leishmaniasis epidemiology and control.

Interactions of Spodoptera littoralis haemocytes following injection with the entomopathogenic fungi: Beauveria bassiana and Nomuraea rileyi.

This study compared the cellular interactions of Spodopteralittoralis haemocytes with two virulence-different entomopathogenic fungi: Beauveriabassiana and Nomuraearileyi. Using light and transmission microscopy, five types of haemocytes namely, prohaemocytes (PRs), plasmatocytes (PLs), granulocytes (GRs), spherule cells (SPs) and oenocytoids (OEs) were identified in the 6th instar larvae. PRs and PLs were found in the haemopoietic tissue. Intra-haemocoelic injection of blastospores induced ultrastructural alterations in the cytoplasm and nuclei of circulating haemocytes of treated larvae. Different responses were observed in the populations of haemocyte types following injection with the tested fungi. The most important changes were the decrease of the numers of GRs accompanied with increase in SPs at 12-48h following injection with B. bassiana, whereas, a decrease of PLs with a commitment increase inSPs and OEs were observed at most time intervals after injection with N. rileyi. Both fungi provoked a decrease of the total number of haemocytes at 48h followed by an increase at 72h post-injection. In vivo assay showed that the GRs and PLs actively phagocytised fungal blastospores. There was a time-dependent decrease and increase in the phagocytosis activity after injection of B. bassiana and N. Rileyi, respectively. In B. bassiana-injected insects, the numbers nodules increased significantly at 6-48h in comparison with the controls post-injection. In N. rileyi-injected insects, nodules increased significantly only at 72h post-injection. No cellular encapsulation was observed in any of the examined insects.

First report of Leishmania tropica from a classical focus of L. major in North-Sinai, Egypt.

Cutaneous leishmaniasis (CL) is prevalent in the Egyptian Sinai Peninsula and previous research has consistently documented the etiologic agent to be Leishmania major. We report the first isolation of Leishmania tropica from human cases of CL in a Northern Sinai community bordering Palestine. Parasite culturing, real-time polymerase chain reaction (PCR), gene sequencing, and restriction fragment length polymorphism (RFLP) analyses indicate CL cases in this community were caused by either L. major or L. tropica (three cases each). Two wild-caught rodents (Gerbillus pyramidum floweri) were infected with L. tropica. Phlebotomus papatasi sand flies were found harboring L. major, however only non-infected individuals of Phlebotomus sergenti, a vector for L. tropica, were caught. Patients with L. tropica had not traveled from the region in over a year, suggesting these cases are autochthonous. This scenario is consistent with an incursion of L. tropica from bordering countries and raises concerns about expansion of this parasite further into Egypt.

Human zoonotic cutaneous leishmaniasis and associated sand flies (Diptera: Psychodidae) in Sheikh Atiya village, southern Sinai, Egypt.

Zoonotic cutaneous leishmaniasis is endemic in Sinai Peninsula. Human cases were recorded from the northern Sinai, however little was known about the disease in southern Sinai. During entomological surveys conducted southern Sinai in summers of 1997-1999, a clinically confirmed ZCL case was encountered for the first time in Sheikh Atiya village in June 1999. The parasite was isolated and identified as L. major using cellulose acetate electrophoresis. A total of 784 phlebotomine sand flies were collected. Sand fly species composition at Sheikh Atiya village showed that P. papatasi and P. alexandri were the most abundant species in the area and each comprises about 47 % of the flies collected. P. sergenti and P. kazeruni occurred in very low numbers. All the female flies dissected (N = 304) were negative for any Leishmania-like flagellates. When the identified isolate was inoculated in the footpads of a clean laboratory colonized Acomys cahirinus dimidiatus, a lesion was developed on the site of inoculation 9 months post injection. The role of P. alexandri and A. c. dimidiatus in transmission of leishmaniasis in southern Sinai remains questionable. Environmental factors contributed to the appearance of ZCL in the area are discussed.