Spatio-temporal patterns of stomoxyine flies (Diptera: Muscidae) in a forested area of Thailand.

Understanding the distribution patterns of vector populations is crucial for comprehending the dynamics of vector-borne diseases. However, data on vector composition and abundance in areas of forest and wildlife-human interface in Thailand remain limited. This research aimed to investigate the spatio-temporal distribution and species diversity of stomoxyine flies (Diptera: Muscidae) in Salakpra Wildlife Sanctuary, Thailand's first wildlife sanctuary. A longitudinal entomological survey was conducted monthly from May 2022 to April 2023 in four habitats: core forest, grassland forest, a wildlife breeding center, and a local cattle farm. A total of 11,256 stomoxyine flies from four genera were captured. Based on morphological keys, nine species of stomoxyine flies were identified: Stomoxys pullus (29.63%), Stomoxys calcitrans (19.65%), Stomoxys indicus (16.09%), Haematostoma austeni (14.23%), Haematobia irritans exigua (8.22%), Haematobosca sanguinolenta (7.96%), Stomoxys uruma (1.98%), Stomoxys sitiens (1.75%), and Stomoxys bengalensis (0.49%). Heterogeneous variations in abundance across months and habitats were observed, in which abundance increased in the rainy season (June-October), exhibiting bimodal peaks at seasonal transitions. Human-disturbed areas, such as the cattle farm, exhibited the highest density and species diversity of stomoxyine flies. In contrast, areas with minimal human disturbance, like core forest, had low diversity and density but supported unique species, like the abundant Haematostoma austeni, which had minor populations in other types of habitats. The results of this study can be integrated into epidemiological models and lay the groundwork for more comprehensive research on vector-borne diseases at the wildlife-livestock interface to mitigate transmission risks and preserve biodiversity.

Title: Schémas spatio-temporels des stomoxes (Diptera : Muscidae) dans une zone forestière de Thaïlande. Abstract: Comprendre les schémas de distribution des populations de vecteurs est essentiel pour comprendre la dynamique des maladies à transmission vectorielle. Cependant, les données sur la composition et l'abondance des vecteurs dans les zones de forêt et d'interface faune-humain en Thaïlande restent limitées. Cette recherche visait à étudier la distribution spatio-temporelle et la diversité des espèces de stomoxes (Diptera : Muscidae) dans le sanctuaire faunique de Salakpra, le premier sanctuaire faunique de Thaïlande. Une enquête entomologique longitudinale a été menée mensuellement de mai 2022 à avril 2023 dans quatre habitats : forêt centrale, forêt de prairie, un centre d'élevage d'animaux sauvages et une ferme d'élevage locale. Au total, 11 256 stomoxes de quatre genres ont été capturés. Sur la base des clés morphologiques, neuf espèces de stomoxes ont été identifiées : Stomoxys pullus (29,63 %), Stomoxys calcitrans (19,65 %), Stomoxys indicus (16,09 %), Haematostoma austeni (14,23 %), Haematobia irritans exigua (8,22 %), Haematobosca sanguinolenta (7,96 %), Stomoxys uruma (1,98 %), Stomoxys sitiens (1,75 %) et Stomoxys bengalensis (0,49 %). Des variations hétérogènes de l'abondance selon les mois et les habitats ont été observées, l'abondance augmentant pendant la saison des pluies (juin à octobre), présentant des pics bimodaux aux transitions saisonnières. Les zones perturbées par l'homme, comme la ferme d'élevage, présentaient la plus forte densité et la plus grande diversité d'espèces de stomoxes. En revanche, les zones peu perturbées par l'homme, comme la forêt centrale, présentaient une faible diversité et une faible densité, mais abritaient des espèces uniques, comme l'abondant Haematostoma austeni, dont les populations étaient mineures dans d'autres types d'habitats. Les résultats de cette étude peuvent être intégrés dans des modèles épidémiologiques et posent les bases d'une recherche plus complète sur les maladies à transmission vectorielle à l'interface faune-bétail afin d'atténuer les risques de transmission et de préserver la biodiversité.

Molecular Identification of Culicoides Species and Host Preference Blood Meal in the African Horse Sickness Outbreak-Affected Area in Hua Hin District, Prachuap Khiri Khan Province, Thailand.

African horse sickness (AHS) was reported as an outbreak in Thailand in 2020. Hematophagous insects from the genus Culicoides are the suspected vector responsible for AHS transmission. Horses in Hua Hin district, Prachuab Khiri Khan province, Thailand, were affected and died from AHS in 2020. However, the potential Culicoides species and its host preference blood meal in the affected areas are unknown. To investigate the potential vectors of AHS, Culicoides were collected using ultraviolet light traps placed near horse stables. Six horse farms, including five farms with AHS history and one farm without AHS history, were included in this study. Morphological and molecular identification of the Culicoides species was performed. Polymerase chain reaction (PCR) targeting the cytochrome b oxidase I (COXI) gene for confirmation of the Culicoides species, identification of the prepronociceptin (PNOC) gene for host preference blood meal, and bidirectional sequencing were conducted. Consequently, 1008 female Culicoides were collected, consisting of 708 and 300 samples captured at positions A and B at a distance of <2 and >5 m from the horse, respectively. Twelve Culicoides species identified by morphology were noted, including C. oxystoma (71.92%), C. imicola (20.44%), C. actoni (2.28%), C. flavipunctatus (1.98%), C. asiana (0.99%), C. peregrinus (0.60%), C. huffi (0.60%), C. brevitarsis (0.40%), C. innoxius (0.30%), C. histrio (0.30%), C. minimus (0.10%), and C. geminus (0.10%). The PCR detection of the Culicoides COXI gene confirmed Culicoides species in 23 DNA samples. PCR targeting the PNOC gene revealed that the Culicoides collected in this study fed on Equus caballus (86.25%), Canis lupus familiaris (6.25%), Sus scrofa (3.75%), and Homo sapiens (3.75%) for their blood meal. Human blood was identified from two samples of C. oxystoma and a sample of C. imicola. Three dominant species including C. oxystoma, C. imicola, and C. actoni that were reported in the Hua Hin area prefer to feed on horse blood. Moreover, C. oxystoma, C. imicola, and C. bravatarsis also feed on canine blood. This study revealed the species of Culicoides in Hua Hin district, Thailand, after the AHS outbreak.

Companion Vector-Borne Pathogens and Associated Risk Factors in Apparently Healthy Pet Animals (Dogs and Cats) in Khukhot City Municipality, Pathum Thani Province, Thailand.

Pet animals (dogs and cats) can be infected with several companion vector-borne pathogens (CVBPs). Morbidity and mortality have been reported in pet animals due to CVBP infections. Pet animals living in close proximity to humans are able to transmit zoonotic pathogens. This study used molecular techniques to investigate the prevalence of CVBPs in apparently healthy pet animals (dogs and cats) from Khukhot City Municipality, Pathum Thani province, Thailand. In total, 210 blood samples were randomly collected from 95 dogs and 115 cats for the detection of seven companion vector-borne pathogens (Anaplasma, Babesia, Bartonella, Ehrlichia, Hepatozoon, Mycoplasma, and Rickettsia) using polymerase chain reaction. The results showed that 10.5% (22/210) of apparently healthy pet animals were infected with at least one pathogen, comprising 6 dogs (6.3% of all dogs tested) and 16 cats (13.9% of all cats tested). Ehrlichia (6.3%) was present only in dogs; furthermore, 1.1% of the dogs were positive for Anaplasma. There was one dog case co-infected with two pathogens (1.1%). In cats, Mycoplasma (9.6%) was the predominant CVBP, followed by Rickettsia (4.4%). The DNA sequences of all positive animals were 97-99% homologous to those found in the GenBank™ database for all CVBPs identified, namely Ehrlichia canis, Anaplasma platys, Rickettsia felis, Mycoplasma haemofelis, and Candidatus Mycoplasma haemominutum. Additionally, the risk of infection with CVBPs in pets was significantly associated with age, with young dogs more likely to be infected with CVBPs than adult dogs (OR 8.5, 95% CI 1.4-50.1, p = 0.006), while adult cats were more likely to be infected with CVBPs than young cats (OR 3.8, 95% CI 1.0-14.0, p = 0.038). The detection of CVBPs demonstrated the potential risk of infection that may occur in apparently healthy pet animals in Pathum Thani province. These results confirmed that apparently healthy pet animals may still be at risk of vector-borne infections and could maintain the infection cycle in pet populations. Furthermore, sampling a greater number of apparently healthy pet animals may disclose predictors of CVBP positivity in domesticated animals in this area.

Preliminary Study on Comparative Efficacy of Four Light Sources for Trapping Culicoides spp. (Diptera: Ceratopogonidae) in Prachuap Khiri Khan Province, Thailand.

The light trap is an important tool to determine the presence and abundance of vectors in the field. However, no one has studied the efficiency of light traps for collecting Culicoides in Thailand. In the present study, the efficacy of four light sources was evaluated in Prachuap Khiri Khan province, Thailand. Incandescent (INCND) light, white fluorescent (WHT-FLR) light, ultraviolet fluorescent (UV-FLR) light, and UV light-emitting diode (UV-LED) light were tested using commercial traps. In total, 30,866 individuals of Culicoides species were collected from November 2020 to June 2021, of which 21,016 were trapped on site 1 and 6,731 were trapped on site 2. The two most abundant Culicoides species were C. imicola (54%) and C. oxystoma (31.2%). UV-FLR was highly effective, followed by UV-LED light, WHT-FLR light, and INCND light, respectively, for Culicoides collection. Significantly, more Culicoides species were collected in those traps baited with UV-FLR light, UV-LED light, or WHT-FLR light than for INCND light traps. Traps equipped with UV-FLR lights can be recommended to trap Culcoides biting midges for monitoring purposes.

Molecular Identification of Cryptosporidium Species from Pet Snakes in Thailand.

Cryptosporidium is an important pathogen causing gastrointestinal disease in snakes and is distributed worldwide. The main objectives of this study were to detect and identify Cryptosporidium species in captive snakes from exotic pet shops and snake farms in Thailand. In total, 165 fecal samples were examined from 8 snake species, boa constrictor (Boa constrictor constrictor), corn snake (Elaphe guttata), ball python (Python regius), milk snake (Lampropeltis triangulum), king snake (Lampropeltis getula), rock python (Python sebae), rainbow boa (Epicrates cenchria), and carpet python (Morelia spilota). Cryptosporidium oocysts were examined using the dimethyl sulfoxide (DMSO)-modified acid-fast staining and a molecular method based on nested-PCR, PCR-RFLP analysis, and sequencing amplification of the SSU rRNA gene. DMSO-modified acid-fast staining revealed the presence of Cryptosporidium oocysts in 12 out of 165 (7.3%) samples, whereas PCR produced positive results in 40 (24.2%) samples. Molecular characterization indicated the presence of Cryptosporidium parvum (mouse genotype) as the most common species in 24 samples (60%) from 5 species of snake followed by Cryptosporidium serpentis in 9 samples (22.5%) from 2 species of snake and Cryptosporidium muris in 3 samples (7.5%) from P. regius.

SPECIES DIVERSITY AND SEASONALITY OF PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) IN SATUN PROVINCE, THAILAND.

Leishmaniasis is prevalent mainly in the southern provinces of Thailand where sand flies are considered to be an important vector. Sand flies were collected using Centers for Disease Control (CDC) light traps in Satun Province from June 2013 to July 2014. A total of 1,982 sand flies (1,228 females and 754 males) were collected. Only female sand flies were identified to the species level and were tested for Leishmania infection using polymerase chain reaction (PCR). Morphological identification revealed 2 genera and 9 species: Phlebotomus stantoni, P. argentipes, Sergentomyia gemmea, S. indica, S. barraudi, S. iyengari, S. bailyi, S. perturbans, and S. silvatica. S. gemmea (57.2%) was the most abundant species. The diversity of sand flies was highest in Thung Wa District. The sand flies were most abundant late in the hot season and early in the rainy season (April to June). The highest number of sand flies was collected in June. Significant correlations between the number of female sand flies and rainfall and between S. gemmea and rainfall were found. Of the female sand flies tested, none were positive for Leishmania spp.

Seasonality and daily flight activity of stable flies (Diptera: Muscidae) on dairy farms in Saraburi Province, Thailand.

Knowledge of seasonal abundance and flight activity patterns are required to design effective management programs for insect pests of humans and livestock. In this study, the seasonality and daily flight activity of Stomoxys species were observed on two dairy farms in Saraburi Province, Thailand. Data were assessed throughout 1 year using Vavoua traps from September 2010 to August 2011. A total of 2,520 individuals belonging to four species were collected. Most Stomoxys species peaked in September (rainy season) and gradually decreased in number toward February (dry season); a second peak occurred between March and April (hot season). Stomoxys calcitrans was caught throughout the year and was the most abundant species in this study. The total number of males and females of S. calcitrans differed significantly among seasons and time intervals. The weather parameters of relative humidity and light intensity were significantly correlated with S. calcitrans abundance.