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: 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.

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: Venom of scorpions are complex bioactive polypeptides. To gain greater insights into the structural and functional impacts of toxins from Androctonus crassicauda (Buthidae), a dangerously venomous scorpion species, its venom was isolated, purified, and characterized.  

Methods: Long chain toxin with four disulfide bonds purified by size exclusion chromatography and reversed-phase HPLC and characterized by amino acid sequencing and molecular weight determination.

Results: The primary structure analysis exhibits a neurotoxin named AnCra2 with 7302.24 Da molecular weight and 64 amino acid residues that cause paralysis and lead to death in NIH mice. The LD₅₀ of AnCra2 was determined to be 0.61±0.04 μg/mice. Phylogenetic analysis displays the toxin has 97% sequence similarity with alpha toxins reported from north African scorpions that affect voltage-gated sodium channels (VGSC), also proposed that differentiation among the scorpions of family Buthidae is affected by the geographical conditions and efficiency in evolutionary varia­tions. AnCra2 exposed binding residues have a high affinity for receptor residues in site-3 (segment-3) of VGSC that are approved by three-dimensional structure and homology modeling.

Conclusion: Purified AnCra2 seems to be a new putative Alpha neurotoxin in homology with the structure of neurotox­ins that act on VGSC as a pharmaceutical tool.

Background: Zoonotic cutaneous leishmaniasis (ZCL) is an endemic disease in Varzaneh City where Leishmania ma­jor is the causative agent and the great gerbil, Rhombomys opimus, is the main reservoir host of the disease. Despite control efforts, ZCL outbreaks recur every few years. This study was conducted to revive information on the parasite/s species circulating between humans and the reservoirs in the region.

Methods: Leishmania infection in patients and R. opimus was studied using direct parasitological and molecular methods during 2019–2021. Nested-PCR and DNA sequencing were used for Leishmania parasite identification. Inter and intra-species variations in the Leishmania parasites were investigated using BLAST and MEGA7 software.

Results: All suspected patients (N=34) and 14 out of 36 great gerbils tested positive for Leishmania parasites via direct parasitological method. Nested-PCR method revealed all the patients were infected with L. major (94.1%) and mixed infection of L. major and Leishmania turanica (5.9%), and great gerbil specimens were infected with either L. major (44.4%), L. turanica (5.6%), or Leishmania gerbilli (5.6%) and also with mixed infection of L. major and L. turanica (30.5%), L. major and L. gerbilli (8.3%) and mix of all the three Leishmania species (5.6%).  

Conclusion: The identical sequences of L. major in both human patients and rodents indicate that the great gerbils are the main reservoirs of L. major in Varzaneh City. The presence of L. turanica in patients would be of interest to carry out further studies to determine the role of this species in the persistence, signs, and treatment of ZCL in humans.

Background: Crimean-Congo hemorrhagic fever (CCHF) is a viral zoonotic disease characterized by high fever, bleeding manifestations, and a high mortality rate. Most patients begin to experience bleeding 5 to 7 days after the onset of the illness, usually while hospitalized. This report highlights that even though patients may develop shock and multi-organ failure, there might be no visible signs of bleeding until shortly before death.

Methods: This case report describes a 42-year-old male patient with a confirmed diagnosis of CCHF via RT-PCR, who died despite intensive care treatment. Despite receiving intensive care treatment, he did not respond to the septic shock therapy and unfortunately passed away within a few hours. The diagnosis of the disease was made by the reference la­boratory with a positive reverse transcriptase-polymerase chain reaction (RT-PCR) test for CCHF.

Results: In this case, the patient exhibited no visible signs of bleeding, such as hematemesis, melena, or gross hematuria, despite being in shock. While CCHF mortality (5–30%) typically occurs during the 3–5-day hemorrhagic phase, this patient died suddenly due to massive intra-abdominal bleeding, skipping the typical bleeding phase.

Conclusion: Clinicians should consider that individual differences may be seen in the clinical course of CCHF disease. In addition, the Coronavirus Disease-2019 (COVID-19) pandemic was experienced worldwide when the patient ap­plied. Given the overlapping initial symptoms of COVID-19 and CCHF, clinicians should prioritize differential diagnosis, es-pecially during the COVID-19 pandemic. During the COVID-19 pandemic, diagnostic delays may occur in the manage-ment of CCHF, particularly due to resource allocation.

Not applicable