Molecular Phylogenetics of Centrocestus formosanus (Digenea: Heterophyidae) Originated from Freshwater Fish from Chiang Mai Province, Thailand

This study aimed to investigate the morphology and reconstruct the phylogenetic relationships of Centrocestus formosanus originating from 5 species of freshwater fish, i.e., Esomus metallicus, Puntius brevis, Anabas testudineus, Parambassis siamensis, and Carassius auratus, in Chiang Mai province, Thailand. Sequence-related amplified polymorphism (SRAP) and phylogeny based on internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 1 (CO1) were performed. The results showed similar morphologies of adult C. formosanus from day 5 after infection in chicks. C. formosanus originated from 4 species of freshwater fish had the same number of circumoral spines on the oral sucker, except for those from C. auratus which revealed 34 circumoral spines. The phylogenetic tree obtained from SRAP profile and the combination of ITS2 and CO1 sequence showed similar results that were correlated with the number of circumoral spines in adult worms. Genetic variability of C. formosanus also occurred in different species of freshwater fish hosts. However, more details of adult worm morphologies and more sensitive genetic markers are needed to confirm the species validity of C. formosanus with 34 circumoral spines originating from C. auratus in the future.

Forensically Important Blow Flies Chrysomya pinguis, C. villeneuvi, and Lucilia porphyrina (Diptera: Calliphoridae) in a Case of Human Remains in Thailand

This is the first study to report Chrysomya pinguis (Walker) and Lucilia porphyrina (Walker) (Diptera: Calliphoridae) as forensically important blow fly species from human cadavers in Thailand, in addition to Chrysomya villeneuvi (Patton) already known in Thailand. In 2016, a fully decomposed body of an unknown adult male was discovered in a high mountainous forest during winter in Chiang Mai province. The remains were infested heavily with thousands of blow fly larvae feeding simultaneously on them. Morphological identification of adults reared from the larvae, and molecular analysis based on sequencing of 1,247 bp partial mitochondrial cytochrome c oxidase subunit 1 gene (CO1) of the larvae and puparia, confirmed the above mentioned 3 species. The approving forensic fly evidence by molecular approach was described for the first time in Thailand. Moreover, neighbor-joining phylogenetic analysis of the CO1 was performed to compare the relatedness of the species, thereby affirming the accuracy of identification. As species of entomofauna varies among cases in different geographic and climatic circumstances, C. pinguis and L. porphyrina were added to the list of Thai forensic entomology caseworks, including colonizers of human remains in open, high mountainous areas during winter. Further research should focus on these 3 species, for which no developmental data are currently available.

Occurrence of Blow Fly Species (Diptera: Calliphoridae) in Phitsanulok Province, Northern Thailand

Based on the current forensic importance of blow flies (Diptera: Calliphoridae), their biological aspects have been studied increasingly worldwide. The blow fly fauna in Phitsanulok Province, Northern Thailand was studied from May 2009 to April 2010 in the residential, agricultural, mountainous and forested areas of Muang, Wat Bot, Nakhon Thai and Wang Thong districts, respectively, in order to know the occurrence of blow flies in this province. Collections were carried out monthly using commercial funnel fly traps and sweeping methods, with 1-day tainted pork viscera as bait. Identification of adult blow flies exhibited 14 634 specimens, comprising of 5 subfamilies, 14 genera and 36 species. Chrysomya megacephala (Fabricius, 1794) and Achoetandrus rufifacies (Macquart, 1843) were the most and second most abundant species trapped, respectively. These two species of carrion flies prevailed in all the types of land investigated. We calculated and compared the diversity indices, species evenness and richness, and similarity coefficients of the blow fly species in various areas. The data from this study may be used to identify the potential of forensicallyimportant fly species within Phitsanulok Province and fulfill the information on blow fly fauna in Thailand.

Effects of human contraceptive on reproduction and offspring in Chrysomya megacephala

OBJECTIVE@#To investigate the effect of human contraceptive (HC) as ability to suppress the reproductive success of blow fly, Chrysomya megacephala (Fabricius) (C. megacephala) and offspring under controlled laboratory conditions.@*METHODS@#Adult C. megacephala were fed with low (0.036 mg/mL) and high dose (0.072 mg/mL) HC (Microgest®, Thailand), containing levonorgestrel and ethinyl estradiol, in their drinking water for 7 days. Three experiments were set; experiment I with fed only in parental males, experiment II with fed only in parental females and experiment III with fed in both males and females. All experiments were then maintained for 3 generations after crossing and inbreeding.@*RESULTS@#A lower ovariole production and less fully mature ovarioles were evident in F1, F2 and F3 than control when parent males, females and both had been fed with high dose HC. Cellular changes during spermatogenesis in F1, F2 and F3 testes was confirmed using transmission electron microscopy (TEM), showing the low level of condensed chromatin, necrotic chromatin, irregularities and degenerated nuclear envelope in the nucleus. In the cytoplasm, mitochondrial swelling, rough endoplasmic reticulum swelling as well as vacuolated cytoplasm were noticed. As for the sperm per se, we found the degenerated nuclei and/or incomplete mitochondrial derivative, axoneme and vacuolated flagella. Regarding deformity in F1, F2 and F3 ovariole, ultrastructural alteration observed by scanning electron microscopy (SEM) included malformations involving fragile enveloping peritoneal sheath, cracked ovarioles, peel away chorion, crumbled eggshell and incomplete development; whereas TEM presented malformed and disorganized mass of cells, proteic yolk granules and vacuolated vesicles.@*CONCLUSIONS@#Administer of HC to adult C. megacephala caused ovariole reduction, less matured ovariole and affected cellular changes in testes and ovariole of offspring up to F3.