Urogenital schistosomiasis, caused by the flatworm-Schistosoma haematobium, is a neglected tropical disease ranked second to malaria in terms of its socio-economic and public health importance in tropical and subtropical areas. Early identification and diagnosis are crucial for effective disease management and control. This study aimed to assess the immunodiagnostic potential of crude Fasciola gigantica-worm antigen (FWA) as a target for detecting anti-S. haematobium antibodies in sera and urine samples. An indirect ELISA was used to assess antibodies against these antigens in serum and urine samples from S. haematobium-infected and non-infected individuals (n=48) in schistosomiasis endemic (NE) and non-endemic (NNE) areas, with microscopy as diagnostic reference. The Area under the receiver operating characteristic (ROC) curve (AUC), sensitivity (SS), and specificity (SP) of FWA for sera in diagnosing S. haematobium-infected individuals using ELISA were 0.957, 97.92%, and 70.82% for positive and NNE samples and 0.542, 70.83%, and 31.25% for infected and NE samples, respectively. Similarly, the diagnostic performances of FWA measured by AUC, SS, and SP were 0.9679, 97.92%, and 83.33% for positive and NNE samples and 0.9670, 97.92%, and 85.33% for positive and NE urine samples, respectively. S. haematobium-specific antibody production against FWA-sera and urine was significantly higher in infected individuals than in non-infected groups in endemic and non-endemic areas (P< 0.0001). This study demonstrated the feasibility of serological diagnosis of urogenital schistosomiasis with Fasciola gigantica worm antigens in resource-constraint where schistosomiasis is endemic. The urine samples exhibited better diagnostic metrics than the sera samples, which can be leveraged as a non-invasive biological sample for diagnostic advantage.

Schistosomiasis remains a major public health concern in endemic regions where rapid and accurate diagnosis is vital for effective control. Persistent diagnostic challenges arise as a result of crude parasite antigen-based assays, which often provoke non-specific immune responses and compromise accuracy. This study validated two immunoreactive proteins eluted from Schistosoma haematobium soluble egg antigen (SEA) and Schistosoma mansoni worm antigen proteins (WAPs) as improved diagnostic tools for field deployment using dot blot assays.

SDS–polyacrylamide gel electrophoresis and Western blot assays identified immunoreactive protein bands from SEA and WAP, which were then evaluated using ELISA on 40 repository samples (20 positive and 20 negative). The bulk of the validation work employed dot blot assays, which is the primary qualitative method used to demonstrate clear immunogenicity of each antigen against patient urine. Statistical analyses, including tests of mean optical density, area under ROC curve (AUC), sensitivity, and specificity, were performed using GraphPad Prism 8.0.1 and MedCalc webserver.

Two highly immunoreactive proteins were successfully eluted: 72 kDa from Sh SEA and 95 kDa from Sm WAP, with diagnostic cut-offs of 0.3383 and 0.1102, respectively. The 72 kDa protein demonstrated excellent performance with 95% sensitivity and 90% specificity, while the 95 kDa protein showed 90% sensitivity and 95% specificity. Both exhibited excellent diagnostic accuracy of 72 kDa (AUC = 0.96; 95% CI: 0.90–1.00) and 95 kDa (AUC = 0.96; 95% CI: 0.91–1.00). Dot blot validation clearly revealed the immunogenic potential of both proteins, visually differentiating positive from negative samples. ELISA results showed significant differences in mean optical density between positive (0.75±0.27), (0.35±0.19) and negative (0.23±0.15), (0.07±0.02) samples (P <0.0001).

The 72 kDa and 95 kDa eluted proteins demonstrated strong immunogenicity and diagnostic precision, addressing the limitations of crude antigen-based assays and offering more specific and reliable tools for schistosomiasis detection in endemic regions.

Ectoparasitic arthropods represent a significant component of faunal biodiversity, with high zoological and veterinary relevance due to their parasitic adaptations and role as vectors of pathogens. This study provides a detailed morphological and biological characterization of four ectoparasitic taxa identified in domestic mammals examined at the “Medicrisvet” Veterinary Clinic (Fălticeni, Suceava, Romania) during 2024: Otodectes cynotis, Rhipicephalus sanguineus, Demodex cati, and Demodex criceti. Specimens were collected from dogs, cats, and hamsters and examined using optical microscopy to document diagnostic morphological traits, developmental stages, and host specificity, with careful attention to subtle variations in body structure, appendages, and cuticular patterns. Quantitative analysis of infestation data revealed Rhipicephalus sanguineus as the dominant species (43% prevalence), followed by O. cynotis (29%), D. cati (18%), and D. criceti (10%), highlighting differences in host preference, seasonal distribution, and potential risk of transmission. The observed morphological variability highlights adaptive features related to host microhabitat, feeding behavior, and environmental interactions. This work contributes new faunistic and morphological data on ectoparasitic arthropods from northeastern Romania and supports their taxonomic documentation within the broader context of veterinary and systematic zoology. The findings emphasize the integrative role of detailed morphological analysis in understanding host–parasite associations, local prevalence patterns, and the regional biodiversity of parasitic arthropods, ultimately providing a basis for improved monitoring and control strategies in domestic animal populations.

Although the skin-scale microstructures of squamate reptiles are known to be highly adapted for habitat-specific functions, including protection, camouflage, and thermoregulation, the structural adaptations of the skin of the color-changing ectotherm Anolis carolinensis have not been systematically investigated. The scale microstructures across key body regions (head, dorsum, venter, tail, and hindleg) of A. carolinensis were meticulously examined using scanning electron microscopy. Statistical analyses were further conducted on the morphological types, density, and distribution of skin sense organs. The scales were predominantly irregular hexagons, although nearly circular scales were observed on the head. Scale area varied significantly across body regions, with the largest scales located on the tail (0.12 ± 0.02 mm²). The head and ventral scales were intermediate in size (0.09 ± 0.04 mm² and 0.09 ± 0.01 mm², respectively), while the dorsal and hindleg scales were the smallest (0.06 ± 0.01 mm² and 0.03 ± 0.01 mm², respectively). The distribution of cutaneous sensory organs was non-uniform across the body. In females, two morphotypes—hair-like and pillow-like sensory organs—were identified, with the former approximately 20 times more numerous. In contrast, male individuals possess only hair-like sensory organs. Moreover, the total number of these organs in males is less than that in females, accounting for approximately half of the total number of sensory organs in females. These organs, located adjacent to a raised median ridge on each scale, were more densely distributed on the head, venter, hindlegs, and tail (0-4 per scale) compared to the dorsum (0-1 per scale). This study details the adaptive microstructures of scales and sensory organs in A. carolinensis, providing key morphological insights into their interactions with the environment. Beyond deepening our knowledge of squamate integumentary adaptations, the work directly contributes to understanding the mechanistic basis of color change and thermoregulation. Furthermore, this study offers valuable insights for the development of novel biomimetic materials, such as adaptive camouflage systems.

Background: The escalating threat of multidrug-resistant (MDR) pathogens underscores the urgent need for novel anti-infective therapeutics. Azadirachta indica (Neem), a traditional medicinal plant, is rich in bioactive limonoids, quercetin derivatives, and azadirachtin with demonstrated antimicrobial activity. However, its metabolomic signature and translational anti-infective potential remain underexplored.

Methods: A combined in vitro and in vivo approach was employed. Methanolic Neem leaf extract was fractionated by LC-MS/MS–based untargeted metabolomics, identifying key metabolites linked to antimicrobial bioactivity. In vitro susceptibility was assessed against MDR Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa using CLSI-standard broth microdilution assays. Synergy with ciprofloxacin and meropenem was evaluated through checkerboard assays. For in vivo validation, BALB/c mice (n = 48) were challenged intraperitoneally with MDR E. coli. Neem extract nanoparticles (50 mg/kg, oral gavage) were administered daily for 10 days, with ciprofloxacin (20 mg/kg) as positive control.

Results: Metabolomic profiling revealed abundant nimbolide, azadirachtin, and quercetin glycosides as major antimicrobial metabolites. Neem extract demonstrated potent in vitro activity with MICs of 64 µg/mL (E. coli), 32 µg/mL (S. aureus), and 128 µg/mL (P. aeruginosa). Checkerboard assays revealed synergistic interactions with ciprofloxacin (FICI = 0.28) and meropenem (FICI = 0.32). In vivo, Neem-treated mice exhibited a 65% survival rate compared to 25% in untreated controls (p < 0.01). Bacterial load in spleen and liver decreased by >1.8 log CFU, with concomitant reduction in serum TNF-α (↓42%) and IL-6 (↓37%).

Conclusion: Neem demonstrates potent anti-infective efficacy through synergistic interactions with antibiotics, immune modulation, and direct antimicrobial activity. Metabolomics-guided identification of active compounds positions Azadirachta indica as a promising medicinal lead for combating MDR infections.

Puberty in sheep represents the activation of the hypothalamus–pituitary–ovary axis, but how estradiol (E2) and its receptors coordinate this transition is still unclear, especially in early-maturing indigenous breeds. Duolang sheep from arid Xinjiang reach first estrus unusually early, offering a natural model to link hormone dynamics, receptor expression, and breed-specific reproductive adaptation across biological levels. Eighteen Duolang ewe lambs were allocated to three stages (prepuberty, puberty, and postpuberty; n = 6 per group). In addition to measuring ERα and ERβ mRNA and protein in the hypothalamus, pituitary, and ovary, we also measured serum E2. Primary ovarian granulosa cells were cultured and exposed to graded E2 concentrations (0–1000 ng/mL) to assess dose-dependent changes in ERα, ERβ, and gonadotropin-releasing hormone (GnRH) expression. Around first estrus, circulating E2 displayed a sharp peak. E2 spiked around first estrus and fell afterward. ERα expression in the hypothalamus and pituitary decreased during puberty and increased post-puberty, indicating a shifting negative feedback role of E2 on the central axis. Conversely, ovarian ERα peaked during puberty and increased gradually from pre- to postpuberty. ERα expression in granulosa cells was biphasic, peaking at 250 ng/mL E2 concentration, while ERβ and GnRH levels increased with higher E2 doses. ERα is the main mediator of E2 feedback in the hypothalamus-pituitary axis, while ERβ is more involved in ovarian maturation and GnRH regulation during pubertal transition. Our study connects molecular receptor dynamics, tissue-level signaling, and breed-level reproductive phenotype, providing candidate biomarkers for selecting high-fertility, early-maturing sheep.

The spotted lanternfly (Lycorma delicatula) is an invasive pest, and its jumping ability is crucial for its dispersal and colonization. While previous studies have reported on adults^’ jumping ability, the differences in jumping capabilities among nymphs in different instars and the underlying biological mechanisms remain unclear. In this study, we employed high-speed videography to quantify jump kinematics (takeoff velocity, acceleration, angle) alongside morphological analysis to examine how jumping performance relates to development across nymphal instars. The results showed that the body lengths of 1st to 4th instar nymphs were 2.59±0.74 mm, 6.31±0.48 mm, 6.85±0.81 mm, 8.28±0.55 mm, and their body weights were 9.78±3.31 mg, 27.94±9.9 mg, 39.13±8.12 mg, 107.37±31.32mg. The nymphs exhibited strong jumping abilities, with takeoff velocities of 0.61±3.42 m/s, 2.17±0.23 m/s, 3.24±0.56 m/s, 4.07±2.31 m/s for 1st to 4th instars. Takeoff velocity increased significantly with instar. The corresponding accelerations were 171.52±0.41g, 89.98±0.67g, 68.32±1.35g, 37.57±2.64g, indicating an inverse relationship between acceleration and body size, which means that smaller nymphs exhibited higher acceleration. Takeoff angles decreased with increasing instars, measuring 171.23±0.43°, 158.31±0.15°, 159.16±0.32°, 155.64±0.13° for 1st to 4th instars. These angles were significantly larger than adult's takeoff angle of approximately 45°. Analysis of the jump trajectory revealed that both vertical and horizontal distances increased with instar. Morphologically, the hind leg lengths of 1st to 4th instar nymphs were 3.43±1.19 mm, 6.63±0.56 mm, 8.11±1.56 mm, 10.30±1.25 mm, and the pleural arch lengths were 1.66±0.16 mm, 2.28±0.19 mm, 3.41±0.89 mm, 3.86±0.52 mm. Analysis indicated that hind leg length and pleural arch length were significantly positively correlated with takeoff velocity, suggesting that the development of these structures is the morphological basis for enhanced jumping performance. This study elucidates the development of jumping ability across nymphal instars and its relationship with biological structural changes, inspiring the design of bioinspired jumping robots.

The dentition of snakes serves a key function in their nutrition, as their feeding strategy is directly related to the shape and size of teeth in various dental bones. The morphology of snake's jaws is highly variable and can be considered an important source of taxonomic information. As such, dentition has been used for over a century to describe and identify taxa, continuing to be an important tool in both taxonomy and phylogeny. The goal of this study was to analyze the maxillary dentition of 15 species of neotropical opisthoglyphous snakes (Clelia plumbea, Helicops carinicaudus, Oxyrhopus clathratus, Oxyrhopus guibei, Oxyrhopus rhombifer, Philodryas aestiva, Philodryas nattereri, Philodryas olfersii, Philodryas patagoniensis, Pseudoboa haasi, Siphlophis compressus, Thamnodynastes nattereri, Thamnodynastes strigatus, Tomodon dorsatus, and Tropidodryas striaticeps), with an emphasis on species found in the state of Paraná, Brazil, as a system to assess intra- and interspecific differences among them. This study aimed to examine the maxillary morphology of snakes with different diets but all belonging to the same colubrid lineage. Additionally, it sought to evaluate the applicability of alternative techniques for osteological material preparation and to interpret the observed variation from functional (diet) and evolutionary perspectives, based on available phylogenies. Two preparation techniques were used: dry preparation and diaphanization methods. The analysis of the specimens was conducted using a Leica stereoscopic microscope, and data obtained were analyzed using PAST software. The results suggested that measuring the jaws alone was insufficient to determine the phylogeny or specialized diet of the assessed opisthoglyphous species.

Background: The N-terminal ectodomain (NTE) of the SARS-CoV-2 membrane (M) glycoprotein is a short, flexible region that remains exposed on the virion surface. Despite its small size and conformational plasticity, this region contains conserved linear epitopes that may serve as practical surrogates for full-length proteins in serological diagnostics.

Objective: The objective of this study was to develop and evaluate a synthetic peptide-based diagnostic assay targeting the NTE of the SARS-CoV-2 M protein.

Methods: Peptide analogs of SARS-CoV-2 M NTE were designed from a cluster of epitopes retrieved on February 2021 from the Immune Epitope Database (IEDB). M1 (CADSNGTITVEELKKLLEQC-am) is a polymerizable peptide used as coating antigen in a diagnostic indirect ELISA, while anti-M1 is an antipeptide antibody affinity-purified from the sera of rabbits immunized with M1i (MADSNGTITVEELKKLLEQC-am). The binding affinity of monomeric and polymeric M1 was estimated in terms of the dissociation constant (K_d), taken from the slope of ELISA data performed against anti-M1, and linearized using the Liliom method. To determine diagnostic accuracy, M1-based indirect ELISA was performed on 221 biobanked plasma controls and on 1,222 serum samples corresponding to 544 RT-PCR–confirmed adult patients admitted to the Philippine General Hospital, a 1,335-bed tertiary referral center, during October 2020–February 2021, before SARS-CoV-2 vaccines became available locally.

Results: Polymeric M1 exhibited a two-fold gain in apparent affinity (K_d = 4.33 nM) compared with the monomeric form (K_d = 8.00 nM). Clinical validation of the M1-based diagnostic ELISA yielded a post hoc (Youden J) sensitivity of 95.26% and specificity of 52.27%, with an overall diagnostic accuracy of 88.70%.

Conclusion: M1 demonstrates that synthetic peptide antigens can serve as stable, high-sensitivity antigens for diagnostic assays. This design principle may be applied to other emerging pathogens where rapid assay development and scalability are critical.

The increasing emphasis on sustainable agricultural practices in the Philippines has intensified the search for eco-friendly and cost-efficient pest control alternatives. Biopesticides, particularly those based on entomopathogenic fungi like B. bassiana, offer a promising solution. This study investigates the enzymatic activity and mass production potential of three native B. bassiana isolates (CD3, CD11, and DSMA1) cultured on readily available agricultural byproducts—banana leaves, corn husks, Pinus kesiya (pine) sawdust, and rice. Enzyme assays measured the isolates' proteolytic, lipolytic, chitinolytic, amylolytic, and ligninolytic activities to assess their substrate compatibility and potential for virulence. Among the three, isolate CD3 exhibited the highest spore yield and was most active in proteolysis and amylolysis, indicating strong degradation and nutrient acquisition abilities. DSMA1 showed the greatest chitinolytic activity, while CD11 was dominant in lipolytic activity. Of the substrates tested, rice supported the highest spore production across all isolates, followed by corn husks and banana leaves. Pine sawdust, however, proved unsuitable due to its high lignin content, which hindered fungal growth. These findings support the use of locally sourced substrates to cultivate effective biocontrol agents. This research contributes to the development of region-specific, sustainable pest management strategies that enhance food security and reduce chemical pesticide reliance in Philippine agriculture.