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An endemic focus of Trypanosoma cruzi infection in a subhuman primate research colony.

Leishmanial forms of Trypanosoma cruzi with multifocal pseudocysts were identified in a deceased adult female rhesus monkey (Macaca mulatta) that had been maintained in an outdoor primate colony in southern Texas. Subsequent clinical investigation of the remainder of the colony identified 20 additional monkeys seropositive for T cruzi. Concurrent epizootiologic surveys confirmed the presence of sylvatic reservoirs and triatomine vectors of T cruzi infection and identified a natural T cruzi transmission cycle. Veterinarians associated with the research colony were informed of the potential for further T cruzi transmission and appropriate insect vector and reservoir animal control programs were implemented..

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Triatomine vectors of Trypanosoma cruzi: a molecular perspective based on nuclear ribosomal DNA markers.

Chagas disease (American trypanosomiasis) is mainly transmitted by blood-sucking bugs of the reduviid subfamily Triatominae (Hemiptera: Prosorrhyncha). Control strategies are directed mainly against these insect vectors, as no vaccine is available and, except in the very early stage of infection, there is no effective chemótherapy. Studies of deoxyribonucleic acid (DNA) will lead to major advances in our knowledge of Triatominae and their relationships to Chagas disease transmission, epidemiology and control. Analyses of complete sequences of nuclear genes coding for ribosomal ribonucleic acid (rRNA) (rRNA genes) and spacers furnish significant information at the levels of higher taxons, genera, species, subspecies, hybrids, varieties and populations of Triatominae. This paper briefly reviews the contributions of studies on the slowly-evolving 18S or small subunit (SSU) rRNA gene and the quickly-evolving second internal transcribed spacer (ITS-2). The whole 18S rRNA gene is a useful marker for supraspecific relationships in Triatominae. ITS-2 is complementary to it, enabling resolution at specific and infraspecific levels. All the evidence suggests that ITS-2 will become the DNA marker of excellence for studies of Triatominae at specific and subspecific levels, as it is in other groups of organisms. Possible applications of data obtained from the study of rRNA and ITS-2 sequences of Triatominae are discussed.

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Trypanosoma rangeli (Tejera, 1920): observations upon pleomorphism.

Meta-trypomastigotes of Trypanosoma rangeli Tejera, 1920, harvested from LIT medium, were inoculated i.p. or s.c. into 6, 16, and 26 g NMRI mice, these representing increasing degrees of immunological maturity. In all cases, similar pleomorphic patterns were observed. Four morphobiometrically differentiable types of trypanosome were encountered in an overlapping temporal sequence. These observations, taken in comparison with those on pleomorphism in this and other species of Trypanosoma by other workers, are consistent with the hypothesis that the pleomorphic types represent the natural development of the parasite, rather than the result of the immune response of the mammal host. Small, slender trypanosomes prevalent at the onset of the parasitemia either reinvade the tissue cells for relatively limited subsequent generations of tissue reproduction, or else differentiate toward the forms that are only capable of colonizing the insect vector.

Age Factors↗

Evaluation of methods for extracting Xylella fastidiosa DNA from the glassy-winged sharpshooter.

The recent spread of the plant pathogenic bacterium Xylclla fastidiosa Wells et al. by an invasive vector species, Homalodisca coagulata Say, in southern California has resulted in new epidemics of Pierce's disease of grapevine. Our goal is to develop an efficient method to detect low titers of X. fastidiosa in H. coagulata that is amenable to large sample sizes for epidemiological studies. Detection of the plant pathogenic bacterium X. fastidiosa in its insect vector is complicated by low titers of bacteria, difficulty in releasing it from the insect mouthparts and foregut, and the presence of substances in the insect that inhibit polymerase chain reaction (PCr). To select the optimal protocol for DNA extraction to be used with PCR, we compared three standard methods and 11 commercially available kits for relative efficiency of X. fastidiosa DNA extraction in the presence of insect tissue. All of the protocols tested were proficient at extracting DNA from pure bacterial culture (1 x 10(5) cells), and all but one protocol successfully extracted sufficient bacterial DNA in the presence of insect tissue. Three DNA extraction techniques, immunomagnetic separation, the DNeasy Tissue kit (Qiagen, Hercules, CA), and Genomic DNA Purification kit (Fermentus, Hanover, MD), were compared more closely using a dilution series of X. fastidiosa (5000-0 cells) with and without insect tissue present. The DNeasy Tissue kit was the best kit tested, allowing detection of 5 x 10(3) X. fastidiosa cells with an insect head background.

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In vivo imaging of malaria parasites--recent advances and future directions.

A new view into the life of malaria parasites is now possible owing to recent advances in imaging techniques and to the generation of tagged parasites. Insights into how parasites interact with their insect vectors and mammalian hosts have been gained by the study of various parasitic forms in their natural environment. Quantitative analysis of Plasmodium ookinete motility has revealed different modes of motility in parasite invasion of the mosquito gut and the extrusion of invaded gut cells from the epithelium. Similar analysis with Plasmodium sporozoites has revealed the importance of parasite motility in transmission from the mosquito vector to the mammalian host.

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Using bacteria to express and display anti-parasite molecules in mosquitoes: current and future strategies.

Vector-borne diseases impose enormous health and economical burdens throughout the world. Unfortunately, as insecticide and drug resistance spread, these burdens will increase unless new control measures are developed. Genetically modifying vectors to be incapable of transmitting parasites is one possible control strategy and much progress has been made towards this goal. Numerous effector molecules have been identified that interfere with parasite development in its insect vectors, and techniques for transforming the vectors with genes encoding these molecules have been established. While the ability to generate refractory vectors is close at hand, a mechanism for replacing a wild vector population with a refractory one remains elusive. This review examines the feasibility of using bacteria to deliver the anti-parasitic effector molecules to wild vector populations. The first half briefly examines paratransgenic approaches currently being tested in both the triatomine bug and tsetse fly. The second half explores the possibility of using midgut bacteria to control malaria transmission by Anopheles mosquitoes.

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[PCR as a tool in confirming the experimental transmission of Leishmania chagasi to hamsters by Lutzomyia longipalpis (Diptera:Psychodidae)].

The use of PCR (polymerase chain reaction) was evaluated for its effectiveness as a tool in the detection of transmission of Leishmania chagasi to a hamster host, Mesocricetus auratus, by insect vector bite. Two pairs of uninfected and anesthetized hamsters were introduced into cages containing infected females of the typical phlebotomine sand fly vector, Lutzomyia longipalpis. The flies were experimentally infected with Leishmania chagasi and the infection was verified by dissection of subsamples. At 37 and 51 days after exposure to the infected flies, biopsies of each hamster's liver and spleen were subjected to direct histopathological and PCR examination. DNA was extracted with Chelex 100; for PCR amplification, primers specific to Leishmania minicircle DNA were used. PCR product was separated on agarose gels and visualized with UV. A band of approximately 120 base pairs was observed in 3 of the 4 biopsies, corresponding to the expected minicircle size. PCR was the only method that detected presence of the parasite. The results demonstrated that the sensitivity of PCR greatly expedites the confirmation process of a particular phlebotomine species as a vector of leishmaniasis.

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Manipulation of wavelength-dependent behaviour of insects: an IPM tool to impede insects and restrict epidemics of insect-borne viruses.

Because of the inability to cure plant virus diseases and the need to protect the environment from toxic pesticides, alternative indirect strategies of disease control are required. In recent decades, virologists have developed non-pesticidal, cultural control practices aimed at reducing the damage caused by these virus diseases by interrupting their epidemiological cycle. Accumulated data on the cues of the host plants and environment associated with visual communication by insect vectors have facilitated the development of cultural practices that interfere with their search, landing and orientation to the crop. This review includes a description of visual cues affecting the phototactic response of insects and cultural practices developed to protect crops from insects and insect-borne viruses by disrupting their searching behaviour and response.

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[Triatominae and Cactaceae: a risk for the transmission of the American trypanosomiasis in the peridomicilary space (Northeast Brazil)].

Field observations carried in semi-arid Brazil Northeast point out the frequent association, in the peridomiciliary space, between a cactus, Cereus jamacaru, the occurrence of nests in its branches and the occurrence of two species of insects vectors of Trypanosoma cruzi, pathogenic agent of Chagas disease: Rhodnius neglectus and Triatoma pseudomaculata. The analysis of the architectural variables of this Cactaceae shows that the presence of nests, and thus of insects, depends on the traditional practices of management of this cactus. This study underlines the relevance of an integrated approach of the ecology of Triatominae for the identification of factors of risk.

Agriculture↗

Epizootiology of bluetongue: the situation in the United States of America.

Bluetongue was first reported in the United States in 1948 in sheep in Texas. The virus has now been isolated from sheep in 19 States. When the disease first occurs in a flock, the morbidity may reach 50 to 75% and mortality 20 to 50%. In subsequent years, the morbidity may be only 1 to 2% with very few deaths. Difference in breed susceptibility has not been observed. Natural bluetongue infection has not been observed in Angora or dairy goats. Bluetongue virus was first isolated from cattle, in Oregon, in 1959. The virus has now been isolated from cattle in 13 States. In cattle, the disease is usually inapparent but can cause mild to severe clinical disease and neonatal losses. Natural clinical bluetongue has also been reported in bighorn sheep, exotic ruminants in a zoo, mule deer, and white-tailed deer. Serological evidence of exposure to the virus has also been found in other species of ruminants in the wild. Inoculation of virulent bluetongue virus, vaccine virus, or natural disease can cause congenital deformities and neonatal losses in calves, lambs, and white-tailed deer fawns. Culicoides is considered the important insect vector of bluetongue. The virus has also been isolated from sheep keds and cattle lice. U.S. field strains of the virus fit into four serologic groups. No cross reactions were found between bluetongue and epizootic haemorrhagic disease of deer viruses. Cattle are considered significant virus reservoirs. It is necessary to use washed erythrocytes, rather than whole blood, and to inoculate susceptible sheep, rather than embryonated chicken eggs, to detect longer-term viraemia in cattle.

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Procyclins, proteases and proteomics: dissecting trypanosomes in the tsetse fly.

The forms of African trypanosomes that live in tsetse fly vectors are coated with lipid-anchored proteins and glycoproteins known collectively as procyclins. Procyclins are expressed during development in the fly in a multiplicity of isoforms yet their functions remain unknown. Recent studies involving a multidisciplinary synthesis of tsetse biology, immunochemistry, biological chemistry and mass spectrometry have yielded much new information about procyclins, which could now provide an unparalleled view of the dynamic molecular interactions between this parasite and its insect vector.

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Bean leaf beetle (Coleoptera: Chrysomelidae) management for reduction of bean pod mottle virus.

Bean pod mottle virus (BPMV) is a management concern for soybean, Glycine max (L.), producers in the North Central states because it can cause yield loss and reduce seed quality by induction of seed coat mottling. The main vector of BPMV is the bean leaf beetle, Cerotoma trifurcata (Forster). An experiment was conducted in 2000 and 2001 at two locations in northwestern and central Iowa to test three insecticide treatments for suppression of bean leaf beetles, and subsequently, BPMV. Treatments of insecticide applications with lambda-cyhalothrin were 1) a single early-season application (23 g [AI] /ha) (2.5 oz/acre) at the VE-VC soybean developmental stage; 2) two early-season applications, the first the same as treatment 1 and a second at the same rate 9-13 d later; 3) a single early-season application the same as treatment 1, followed by a mid-season application (28 g [AI] /ha (3.2 oz/acre) at approximately R2 (flowering, near 15 July); and 4) an unsprayed control. Application of lambda-cyhalothrin after soybean emergence and again as first-generation bean leaf beetles emerged in northwestern Iowa in 2000 (treatment 3) significantly reduced beetle densities through mid-season, BPMV field incidence by 31.5%, and seed coat mottling by 31.2%, compared with the unsprayed control. Similar effects were measured at the same location when insecticide was applied twice at early season (treatment 2). Yield was 453.7 kg/ha (6.74 bu/acre) greater in treatment 2 and 525.20 kg/ha (7.80 bu/acre) greater in treatment 3 than in the unsprayed control at the northwestern site in 2000. At both locations in 2001 fewer treatment effects were observed, which was likely related to lower beetle populations in that year. Early-season insecticide sprays targeted at overwintered beetles on VC-VE reduced the initial population of vector insects and may have contributed to a lower first-generation population because of reduced overwintered beetle oviposition. In 1 year at one location there was a benefit to an additional mid-season insecticide spray, although effectiveness of spraying at this time could vary based on the magnitude of the vector population.

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Maize and oat antixenosis and antibiosis against Delphacodes kuscheli (Homoptera: Delphacidae), vector of "Mal de Rio Cuarto" of maize in Argentina.

"Mal de Rio Cuarto" (MRC) is the most important virus disease of maize, Zea mays L., in Argentina. Several maize lines show different levels of resistance to MRC in the field; however, no studies have been conducted to investigate resistance mechanisms against its insect vector, Delphacodes kuscheli Fennah (Homoptera: Delphacidae). Oat, Avena spp., is the main overwintering host of D. kuscheli and main source of populations that infest maize. Although oat varieties resistant to the greenbug, Schizaphis graminum (Rondani) (Homoptera: Aphididae) are commercially available, their effect on D. kuscheli is unknown. We conducted laboratory experiments to test for the presence of antixenosis and antibiosis resistance mechanisms on six maize lines with different levels of field resistance to MRC, and seven commercial oat cultivars that include two S. graminum-resistant varieties. We did not find antibiotic effects of maize lines on D. kuscheli longevity and survivorship patterns, but we obtained antixenotic effects from the LP2 line (field moderate) due to reduced settling preference and feeding. Oat 'Bonaerense Payé and 'Suregrain INTA' showed both antixenosis and antibiosis, with significantly less settling preference, oviposition in the no-choice test, and reduced total fecundity in comparison with the other varieties studied. The S. graminum-resistant 'Boyera F. A.' and 'Tambera F. A.' did not showed a consistent pattern of resistance versus D. kuscheli across all experiments. Our results indicate the presence of potential sources of insect resistance in the maize lines and oat cultivars tested that may be used in MRC integrated pest management programs.

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Population dynamics of Plasmodium sporogony.

Malaria transmission relies on the sporogonic development of Plasmodium parasites within insect vectors. Sporogony is a complex process that involves several morphologically distinct life-stages and can be described in terms of population dynamics: changes in the abundance and distribution of successive life-stages throughout development. Recent publications on the population dynamics of sporogony are reviewed, with special attention to the differences and similarities among the parasite-vector systems examined thus far. Understanding the population dynamics of malaria parasites within their natural vectors will lead to a better understanding of how malaria parasites survive and are maintained within mosquitoes.

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Black fly involvement in the epidemic transmission of vesicular stomatitis New Jersey virus (Rhabdoviridae: Vesiculovirus).

The transmission routes of Vesicular stomatitis New Jersey virus (VSNJV), a causative agent of vesicular stomatitis, an Office International des Epizooties List-A disease, are not completely understood. Epidemiological and entomological studies conducted during the sporadic epidemics in the western United States have identified potential virus transmission routes involving insect vectors and animal-to-animal contact. In the present study we experimentally tested the previously proposed transmission routes which were primarily based on field observations. Results obtained provide strong evidence for the following: (1) hematophagous insects acquire VSNJV by unconventional routes while blood feeding on livestock, (2) clinical course of VSNJV infection in livestock following transmission by an infected insect is related to insect bite site, (3) infection of livestock via insect bite can result in multiple transmission possibilities, including animal-to-animal contact. Taken together, these data significantly add to our understanding of the transmission routes of a causative agent of one of the oldest known infectious diseases of livestock, for which the details have remained largely unknown despite decades of research.

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Insect-mediated transmission of mixed and reassorted cucumovirus genomic RNAs.

Transmissions of virus using the aphid Myzus persicae were performed using plants co-infected with two cucumoviruses, tomato aspermy virus (V-TAV) and cucumber mosaic virus (M-CMV). Five of the aphid-transmitted progeny viruses (3.7%) induced symptoms distinct from those induced by either parental virus. Northern blot hybridization analysis of encapsidated RNAs from these novel progeny demonstrated that all of the RNA profiles were characteristic of pseudo-recombinants, i.e. viruses with reassorted genomic RNAs. The two larger RNAs, 1 and 2, originated from V-TAV, whereas RNA 3 was derived from M-CMV. A more sensitive RNase protection assay analysis of both unencapsidated and encapsidated RNAs revealed the presence of minor populations of V-TAV-derived RNA 3 in all of these novel progeny, and of M-CMV-derived RNA 1 (and presumably RNA 2) in one of the progeny. A bias against the encapsidation of the minor populations of RNAs by the M-CMV coat protein was observed, suggesting that there is specificity or competition with regard to the encapsidation of cucumoviral RNAs in vivo. This study demonstrates that insect vectors can mediate the establishment of pseudorecombinants with mixed populations of RNA 3.

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VSV-NJ on Ossabaw Island, Georgia. The truth is out there.

Ossabaw Island, Georgia, is the only recognized enzootic focus of vesicular stomatitis virus New Jersey (VSV-NJ) in the United States and has been the subject of VSV-NJ research since 1981. To date, VSV-NJ antibodies have been detected only from feral swine, cattle, equines, deer, and raccoons. VSV-NJ transmission occurs annually, is seasonal, and is associated with the maritime forest. Despite high transmission rates the clinical disease is rarely detected. A sand fly (Lutzomyia shannoni) occurs on the Island, and experimental and field data suggest that it is a biological vector of VSV-NJ at this site. Many questions relating to the epidemiology of VSV-NJ on Ossabaw remain. What is the maintenance cycle of VSV-NJ? Is a vertebrate amplifying host(s) needed? Are other insect vectors involved in mechanical or biological transmission? Why do vesicular lesions develop on some but not all infected animals? Do native and domestic animals play the same role in the maintenance cycle? These questions challenge researchers in all areas where VSV-NJ occurs. It is our hope that Ossabaw Island will provide a much needed model system for gaining insight into the epidemiology of this virus.

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