On the true identity of Sergentomyia gemmea and description of a closely related species: Se. raynali n. sp.

Several species of Leishmania are responsible for leishmaniases in Thailand, although little is known about their transmission. Sergentomyia gemmea has been suspected several times to transmit Leishmania martiniquensis. Some captures carried out in Thailand and Lao People's Democratic Republic have emphasized the scarcity of Se. gemmea, comprising only 1% of the collected females. The sequencing of cytochrome B mtDNA of our specimens showed that our specimens are not grouped with other Se. gemmea previously deposited in GenBank. The latter are grouped with some Se. khawi and Se. hivernus that we processed in the present study. We suspect misidentifications and propose focusing on the most useful characters for identification of Se. gemmea based on the examination of type-specimens. The examination of the ascoids exhibiting anterior spurs is the most important one. However, we also describe Se. raynali n. sp. exhibiting comparable spurs but differing from Se. gemmea by its original cibarium. Finally, the vectorial role of Se. gemmea appears very questionable in the absence of new evidence.

Species composition and population dynamics of phlebotomine sand flies in a Leishmania infected area of Chiang Mai, Thailand.

Phlebotomine sand flies are established vectors of leishmaniasis in humans. In Thailand, Leishmania martiniquensis and "Leishmania siamensis" have been described as causative agents of leishmaniasis. In this study, a survey of sand flies in the Leishmania infected area of Hang Dong district, Chiang Mai, Thailand was performed using CDC light traps for eight consecutive months, from January to August 2016. A total of 661 sand flies were collected, and of 280 female sand flies, four species of the genus Sergentomyia including Sergentomyia gemmea, S. barraudi, S. indica, and S. hivernus and one species of the genus Phlebotomus, Phlebotomus stantoni, were identified. S. gemmea and S. hivernus were found in Chiang Mai for the first time. The density of captured female sand flies was high in warm and humid periods from June to August, with temperatures of around 26°C and relative humidity about 74%. In addition, S. gemmea was the most predominant species in the area. Further studies as to whether or not these sand fly species could be a vector of Leishmaniasis in Thailand are required.

Didilia sp. infecting Phlebotomus stantoni in Thailand.

Nematode infection in wild caught Phlebotomine sand flies was investigated in Thailand. Light microscopy (LM) and scanning electron microscopy (SEM) were used to detect and morphologically characterize entomopathogenic nematodes that presented in the sand flies. Didilia sp. nematodes were found for the first time in the body cavity of wild caught male Phlebotomus stantoni sand flies. The Didilia sp. was identified based on the morphology of the adult nematodes, from their stylet and teeth at the anterior tip, body length, and egg shell sculpture. It was noted that every infected male sand fly had unrotated genitalia, which would not allow them to mate, thus leading to the loss of their offspring. This finding provided information that might lead to study on whether or not the Didilia sp. has the potential to control sand fly population.

Detection of Leishmania martiniquensis DNA in various clinical samples by quantitative PCR.

Leishmaniasis is a neglected tropical parasitic disease affecting a large number of countries in the world. Early diagnosis of Leishmania infections is essential for therapeutic reasons, as it can decrease morbidity and mortality. L. siamensis and L. martiniquensis are novel Leishmania species recently described in Thailand and Myanmar. The disease is usually found in immunocompromised patients, especially those who have AIDS. Currently, the diagnosis of Leishmania infection in Thailand relies on microscopy, microbial culture, and polymerase chain reaction (PCR). In this study, we established a quantitative PCR (qPCR) method for detection of L. martiniquensis DNA in various types of clinical specimens, including whole blood, buffy coat, saliva, and urine of L. martiniquensis infected patients. The results of the qPCR assay were positive in all saliva samples. The assay is therefore effective to detect L. martiniquensis DNA even in noninvasive specimens, and it could be used for the diagnosis, follow up, and survey of L. martiniquensis infections.

Morphological and protein analyses of adult female salivary glands of Anopheles barbirostris species A1 (Diptera: Culicidae).

Morphology and protein profiles of female salivary glands of Anopheles barbirostris species A1 were analyzed. Female glands consisted of a distinctive tri-lobed structure connected to a main salivary canal, a single medial and two lateral lobes with proximal and distal portions. Cellular architecture was similar among the lobes, with secretory material appearing as large masses. Cells of the proximal-lateral lobes contained secretory masses with a finely filamentous aspect. In the distal-lateral lobes, cells had a dense secretory product with mottled pattern. Cells of the medial lobe had secretory masses which were uniformly stained and highly electron dense. Following emergence, the glands accumulated secretory material rapidly and developed completely within three days. Degenerative changes including loss of stored secretion and increase of cytoplasmic vacuolation and concentric lamellar structures were observed from day 16 post emergence that correlated with total amount of the salivary gland proteins determined during development. SDS-PAGE, nanoLC-MS, and glycoprotein analysis revealed at least eleven major protein bands, of which each morphological region contained different major proteins. Two glycoproteins, apyrase/5'-nucleotidase and D7, were identified. These results form a basis for further studies on details of cytopathological changes of malarial infected glands and roles of the proteins in disease transmission.

First outbreak of human trichinellosis caused by Trichinella pseudospiralis.

This is the first report of an epidemic of human infection with Trichinella pseudospiralis. An outbreak of trichinellosis affecting 59 individuals, of whom one died, occurred in southern Thailand during 1994-1995. The source of this epidemic was raw pork from a wild pig that was distributed to villagers by a local hunter. The most striking clinical features among 50 individuals who could be followed were muscular swelling, myalgia, and asthenia persisting for > 4 months. These were associated with significant elevations of creatine phosphokinase and lactate dehydrogenase levels. All patients had Trichinella-specific IgG antibodies in an enzyme-linked immunosorbent assay. Muscle biopsies, performed in six cases, showed nonencapsulated, actively migrating Trichinella larvae. Experimental infection of mice with larvae from human biopsies revealed nonencapsulated muscle larvae consistent with T. pseudospiralis. The identification of muscle larvae from a human specimen by random amplified polymorphic DNA analysis confirmed the causative agent to be T. pseudospiralis. Patients seemed to respond best to treatment with albendazole.