Background: Fleas are blood-sucking ectoparasites with complete metamorphosis. They belong to the order Si­phonaptera and can infest both humans and animals, causing dermatitis and transmitting vector-borne diseases. De­spite extensive study of their classification and biology, the phylogenetic relationship between fleas in Iran is not fully understood. This research aimed to identify the flea species collected from different parts of East Azerbaijan Province in northwest Iran, using morphological, molecular, and phylogenetic analysis.

Methods: From October 2019 to October 2020, we collected fleas using various methods such as hand catching for humans, brushing for dogs and cats, sticky traps for rodent burrows, light traps, and dishes with water for sheep and goats. After identifying the flea species using morphological identification keys, we extracted total genomic DNA and amplified it by targeting the ITS2 region. The PCR products were then directly sequenced to investigate the flea species.

Results: In total, 1929 flea specimens were collected, revealing three genera and four species. The breakdown of the specimens is as follows: Pulex irritans (n=1206; 62.5%), Ctenocephalides canis (n=345; 18%), Ctenocephalides felis felis (n=203; 10.5%), Ctenocephalides felis orientis (n=160; 8%), and Xenopsylla nuttalli (n=15; 1%). Phyloge­netic analysis indicated low to moderate haplotype diversity (Hd: 0−0.524) across five distinct clades: P. irritans, C. canis, C. felis felis, C. felis orientis, and X. nuttalli.

Conclusion: This study represents the first in-depth analysis in East Azerbaijan Province, highlighting the signifi­cance of considering P. irritans as a major vector when assessing the risk of local disease transmission.

Background: One of the main global public health issues that affect several facets of the population is malaria.  Ethio­pia primarily uses Indoor Residual Spraying (IRS) and Long-lasting Insecticidal Nets (LLINs) as lifesaving frontline malaria vector prevention and control interventions. This study intends to assess the trend of malaria prevalence and coverage and utilization of IRS and LLINs in Itang district, western Ethiopia.

Methods: A retrospective analysis was conducted on malaria cases reported by Health Centers from 2017 to 2021. Data were gathered between July 02, 2021, and October 30, 2021, using a community-based cross-sectional household sur­vey to evaluate the coverage and utilization of IRS and LLINs. The chi-square test was used to compare the trend of malaria prevalence among sex and age groups. Statistical significance was considered at P< 0.05.

Results: Among 4,500 patients with suspected cases of malaria, 40% of the cases had microscopically confirmed malar­ia. Plasmodium falciparum was more prevalent than P. vivax in the district. Only 368 (38.25%) people slept under the net yesterday night out of the 962 people who utilized it overall during the study period. 127 (63.5%) households re­ported no insecticide spraying in the past 12 months, while 73 (36.5%) reported their houses were sprayed.

Conclusion: The number of malaria cases reached its peak in 2019 and hit its lowest point in 2021.  Results show that IRS and LLINs coverage and utilization were poor and warrant community awareness and support by all concerned bodies.

Background: Anaplasma spp. and Ehrlichia spp. are amongst the most important tick-transmitted bacteria that can cause zoonotic disease in various hosts including ruminants and humans.

Methods: In this study, 16srRNA, EE, and dsb sequences were respectively used to screen Anaplasmataceae family, Anaplasma spp., and Ehrlichia spp. in tick samples (n= 100) collected from 100 domestic ruminants including 50 sheep and 50 cattle in Jiroft City, southeast of Iran, between June and August 2021.

Results: two genera were predominant among the ticks including Hyalomma spp. (64%; 43% from sheep and 21% from cattle) and Rhipicephalus spp. (36%; 22% from cattle and 14% from sheep); all ticks were adult and 73% of them were male. DNA of Anaplasmataceae was detected in 17% (17/100) of the ticks collected from cattle (18%; 9/50) and sheep (16%; 8/50). Anaplasma spp. was not found in the samples, but two ticks were positive for Ehrlichia spp.; all were posi­tive for Ehrlichia spp. belonged to the cattle (4%; 2/50).

Conclusion: This study shows that Anaplasmataceae strains are circulating via ticks among domestic ruminants in the study area, emphasizing the need for effective tick control strategies by livestock farmers, health, and veterinary au­thorities. Surveillance, molecular characterization and further sequencing-based studies are crucial for informed control and prevention efforts.

Background: Visceral leishmaniasis, commonly known as kala-azar, and prevalent in more than 70 countries and sev­eral regions of Iran. It is one of the main diseases transmitted by sand flies.

In this work, geometric morphometrics and DNA barcoding were employed as novel techniques to enhance the diagnos­tic tools used in this study.

Methods: Phlebotomus perfiliewi transcaucasicus, Phlebotomus major s.l., and Phlebotomus tobbi caught from three districts in the Ardabil Province, northwest of Iran. The right wings of 286 female sand flies were analyzed using geo­metric morphometric (GM) tools. Additionally, the COI gene was isolated from each of the three species, amplified using universal primers, and sequenced through the DNA barcoding method for classification. This sequencing data was then formatted to generate morphometric analyses.

Results: The landmarks with the most variations were found in sets 10, 12, 13, and 14, whereas the first set's landmarks at 1 and 11, along with those from the second set at positions 2, 3, and 5 exhibited the greatest variations. Analysis of the size and shape variations in the wings indicates the presence of distinct populations (P< 0.05). Furthermore, the DNA barcoding results not only confirmed the findings from the geometric morphometric analysis but also revealed both interspecific and intraspecific distances.

Conclusion: This study was the first attempt to assess whether wing geometry morphometrics, combined with DNA barcode techniques, can effectively distinguish the three mentioned species in the studied areas. Furthermore, the identi­fication of Phlebotomus neglectus in this area prompted recommendations for additional research.

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