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Cryptosporidium is one of the most important parasitic protozoa that can be transmitted through food and water contamination. With the increasing report of Cryptosporidium infections in wild birds, especially in herbivorous waterfowl, concerns have been raised for oocyst contamination of water and food supplies, which in turn can cause human and domestic animal infections in areas neighboring wild birds’ habitats. This review discusses the epidemiology, species, and genotypes distribution of Cryptosporidium in wild birds around the world. The overall prevalence of Cryptosporidium in wild birds was calculated as 3.96% (1945/49129), with 6 Cryptosporidium species (C. andersoni, C. parvum, C. meleagridis, C. avium, C. baileyi, and C. galli) and 5 genotypes (Goose genotype I, Goose genotype II, Avian genotype I, Avian genotype III, and Avian genotype VI) reported. As wild birds mainly live in the wild, control method for the Cryptosporidium infection in wild birds is still lacking, which increases the probability of disease transmission from wild birds to humans. The main purpose of this review is to highlight the Cryptosporidium infection in wild birds and its transmission, associated risk factors, and their prevention, illustrating the necessity of multidisciplinary approaches toward screening and control of Cryptosporidium infections.

Background Cryptosporidium parvum is an enteric protozoan parasite with zoonotic importance and can cause cryptosporidiosis in humans as well as domestic and wild animals worldwide. The IId subtype family (SF) is one of the most prevalent subtypes of C. parvum . Some clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) protein systems have been developed to detect nucleic acid with high flexibility, sensitivity and specificity. Methods By integrating recombinase polymerase amplification and the C as12a/crRNA trans - c leavage system (termed ReCTC), we established end-point diagnostics by observing fluorescence readouts with the naked eye under blue light and on-site diagnostics using a lateral flow strip (LFS) biosensor. Results Our ReCTC-based diagnoses can detect as little as a single copy of a cloned C. parvum 60-kDa glycoprotein (GP60) gene, 10 oocysts per gram (OPG), clinical fecal sample without tedious extraction of genomic DNA and have no cross-reactivity with other SFs of C. parvum or other common enteric parasitic protozoa. Conclusions This study provided a new strategy for direct identification of the IId SF of C. parvum free of highly trained operators and expensive special equipment. Graphical Abstract

This study focused on the fast vehicle route optimization issue with carbon emission and time window constraints for on-time consumer demand based on the greedy approach. A greedy algorithm was established to rapidly plan the distribution route to obtain the lowest distribution cost and shortest distribution time to achieve a green distribution. The distribution cost model covering the costs of vehicle transportation time, time window deviation, fossil consumption, and PM2.5 emission was established based on on-time demand and green distribution characteristics. This study analyzed the milk distribution route for Guangxi A Diary Co., Ltd. based on the greedy algorithm. The results show that compared with the genetic algorithm, the algorithm running time is reduced by 2 s, although the greedy algorithm requires an additional cost of 21.73 CNY.

Background Cubital tunnel syndrome (CuTS) is a commonly seen form of compressive neuropathy. This study aimed to assess the clinical value of 3D-FIESTA-C for evaluating the arcuate ligament in patients with CuTS. Methods Between October 2018 and May 2019, 22 inpatients diagnosed with CuTS were enrolled (15 males, 7 females; mean age 60.91 ± 7.38 years). All patients underwent preoperative 3D-FIESTA-C scanning of the elbow joint and subsequently received cubital tunnel release. The accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and kappa value of the 3D-FIESTA-C sequence in displaying the arcuate ligament in its entirety and ulnar nerve were calculated using the intraoperative results as the gold standard, and the consistency test between this sequence and intraoperative arcuate ligament was compared. Paired t-test was used to compare the differences of arcuate ligament thickness measured by the 3D-FIESTA-C sequence and intraoperative exploration. P  < 0.05 was considered statistically significant. Results The 3D-FIESTA-C revealed a comparatively low display rate on the site of olecranon and olecranon attachment when evaluating arcuate ligament (81.8% and 68.2%, respectively). Intraoperative exploration showed the absence of arcuate ligament and ulnar nerve dislocation in 3 patients, while the 3D-FIESTA-C revealed the absence of the arcuate ligament in 2 patients. Using ulnar neurolysis of the cubital tunnel as the gold standard, the accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and Kappa of the 3D-FIESTA-C for displaying the arcuate ligament in its entirety were 81.8%, 79.0%, 100%, 100%, 42.9%, and 0.51, respectively. When using the 3D-FIESTA-C to display the arcuate ligament on the site of the ulnar nerve, the above values were 95.5%, 100%, 66.7%, 95.0%, 100%, and 0.78, respectively. There was no significant difference in arcuate ligament thickness measured by the 3D-FIESTA-C and intraoperative exploration (0.58 ± 0.15 mm Vs. 0.68 ± 0.34 mm, P  = 0.37). Conclusions The 3D-FIESTA-C can accurately display the morphological features and positions of the arcuate ligament of the elbow joint and its relationship with the ulnar nerve. And it can also be used to conduct quantitative measurement of the thickness of the arcuate ligament. Clinical trial number Not applicable.

Objective Intervertebral disc degeneration (IVDD) has been closely associated with ferroptosis in nucleus pulposus cells (NPCs), the underlying regulatory mechanisms and therapeutic strategies remain poorly defined. This study aims to delineate how ginsenoside Rg3 mitigates IVDD progression through ferroptosis suppression, providing a basis for clinical translation. Method An erastin‐induced nucleus pulposus cell ferroptosis model was established. Suitable Erastin concentrations (0–20 μM) were screened via CCK‐8, qRT‐PCR, and Western blotting based on viability, extracellular matrix (COL2A1/ACAN/ADAMTS5/MMP3) and ferroptosis markers (GPX4/FTH‐1/ACSL4), followed by determination of optimal Rg3 concentrations (0–150 μM) using identical methods. Key targets of Rg3 were predicted through network pharmacology and verified by qRT‐PCR and Western blotting. After establishing a rat tail puncture‐induced IVDD model, local injection of Rg3 was administered. Therapeutic efficacy was evaluated by MRI assessment of nucleus pulposus status and disc height, alongside histological and immunohistochemical analyses of Rg3's role in delaying disc degeneration. Result 5 μM Erastin effectively induced ferroptosis in nucleus pulposus cells, reducing cell viability, suppressing expression of extracellular matrix anabolic proteins (COL2A1, ACAN), while promoting catabolic factors (MMP3, ADAMTS5) and downregulating ferroptosis inhibitors (GPX4, FTH‐1). These alterations were significantly reversed by 100 μM Rg3. Integrated network pharmacology and molecular biological validation identified PRKAA2 as the key target mediating Rg3's anti‐degenerative effects. In vivo rat experiments demonstrated that Rg3 treatment preserved disc height and attenuated disc degeneration, with histological and immunohistochemical analyses further confirming its therapeutic efficacy and PRKAA2‐targeted regulation. Conclusion This study elucidates the therapeutic mechanism of Rg3 in delaying IVDD progression via PRKAA2‐mediated ferroptosis inhibition, providing substantial experimental evidence for its clinical translation as a potential disease‐modifying agent. Ginsenosides Rg3 were used to repair the degenerative nucleus pulposus; Furthermore, Rg3 was identified as the bioactive molecule that inhibits the occurrence of ferroptosis through inhibiting PRKAA2 expression in degenerated discs.

Background Captive wild animals in zoos infected with Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi , and Blastocystis sp. can be sources of zoonotic infections and diseases. Therefore, to investigate the distribution of these pathogens in captive wild animals of zoos in Henan, China, a total of 429 fresh fecal samples were collected from six zoos in Henan, China. The infection rates of Cryptosporidium spp., G. duodenalis , E. bieneusi , and Blastocystis sp. were determined by PCR analysis of corresponding loci. Positive results for Cryptosporidium ( C. parvum and C. hominis ) were subtyped based on the ( gp60 ) gene. Results The overall prevalence was 43.1% (185/429), and the prevalence of Cryptosporidium , Giardia duodenalis, Enterocytozoon bieneusi , and Blastocystis sp. were 2.8% (12/429), 0.5% (2/429), 20.8% (89/429), and 19.1% (82/429), respectively. Five Cryptosporidium species, namely, C. hominis , C. parvum , C. muris , C. andersoni , and C. macropodum , were identified in this study. Cryptosporidium parvum was further subtyped as IIdA19G1. Two Giardia duodenalis assemblages (A and E) were also identified. A total of 20 Enterocytozoon bieneusi genotypes were detected, including 18 known (BEB6, D, HND-1, CD7, SDD1, Henan-IV, KIN-1, CHK1, Peru8, Henan-V, CHG11, CHG-1, CHS9, CHG21, Type-IV, CHC9, CM5, and CHB1) and 2 novel genotypes (CHWD1 and CHPM1). A total of nine subtypes of Blastocystis sp. (ST1, ST2, ST3, ST5, ST6, ST7, ST10, ST13, and ST14) were identified in captive wild animals in zoos in the present study. Cryptosporidium andersoni, nine Enterocytozoon bieneusi genotypes, and five Blastocystis subtypes were here first identified in new hosts. Conclusions Our study has expanded the host ranges of these four pathogens. The data indicate that animals in zoos can commonly be infected with these four zoonotic pathogens, and animals in zoos are potential sources of zoonotic infections in humans.

Background With the promotion of “One Health,” the health of animals and their impact on the environment have become major concerns recently. Widely distributed in China, the whooper swans ( Cygnus cygnus ) and black swans ( Cygnus atratus ) are not only important to the ecological environment, but they may also potentially influence public health security. The metagenomic approach was adopted to uncover the impacts of the gut microbiota of swans on host and public health. Results In this study, the intestinal microbiome and resistome of migratory whooper swans and captive-bred black swans were identified. The results revealed similar gut microbes and functional compositions in whooper and black swans. Interestingly, different bacteria and probiotics were enriched by overwintering whooper swans. We also found that Acinetobacter and Escherichia were significantly enriched in early wintering period swans and that clinically important pathogens were more abundant in black swans. Whooper swans and black swans are potential reservoirs of antibiotic resistance genes (ARGs) and novel ARGs, and the abundance of novel ARGs in whooper swans was significantly higher than that in black swans. Metagenomic assembly–based host tracking revealed that most ARG-carrying contigs originated from Proteobacteria (mainly Gammaproteobacteria ). Conclusions The results revealed spatiotemporal changes in microbiome and resistome in swans, providing a reference for safeguarding public health security and preventing animal epidemics.

The complex and variable sea ice environment constrains the safety and efficiency of Arctic shipping. To address the limitations of existing methods in balancing long-term planning with dynamic route adjustment ahead of the vessel, this paper develops an Arctic route decision-making system based on strategic–tactical coordination. The system establishes a “Global Guidance–Local Correction” collaborative mechanism: the strategic module utilizes medium-range climate forecast data to generate a set of daily baseline routes over a 30-day forecast window, providing macro-scale route guidance and feasibility assessment for the voyage; meanwhile, the tactical module integrates high-spatiotemporal-resolution sea ice concentration (SIC) data to screen local high-SIC areas that constrain navigation and execute tactical route correction ahead of the vessel. Experimental results demonstrate the feasibility of integrating long-term global planning with dynamic route adjustment. This provides a feasible route planning framework for integrating long-term strategic guidance with tactical-level dynamic adjustment under complex Arctic ice conditions, offering technical support for future intelligent Arctic navigation systems.

Background Eimeria are obligate protozoan parasites, and more than 1,500 species have been reported. However, Eimeria genomes lag behind many other eukaryotes since obtaining many oocysts is difficult due to a lack of sustainable in vitro culture, highly repetitive sequences, and mixed species infections. To address this challenge, we used whole-genome amplification of a single oocyst followed by long-read sequencing and obtained a chromosome-level genome of Eimeria tenella . Results The assembled genome was 52.13 Mb long, encompassing 15 chromosomes and 46.94% repeat sequences. In total, 7,296 protein-coding genes were predicted, exhibiting high completeness, with 92.00% single-copy BUSCO genes. To the best of our knowledge, this is the first chromosome-level assembly of E. tenella using a combination of single-oocyst whole-genome amplification and long-read sequencing. Comparative genomic and transcriptome analyses confirmed evolutionary relationship and supported estimates of divergence time of apicomplexan parasites and identified AP2 and Myb gene families that may play indispensable roles in regulating the growth and development of E. tenella . Conclusion This high-quality genome assembly and the established sequencing strategy provide valuable community resources for comparative genomic and evolutionary analyses of the Eimeria clade. Additionally, our study also provides a valuable resource for exploring the roles of AP2 and Myb transcription factor genes in regulating the development of Eimeria parasites.

Brucella melitensis is a significant zoonotic pathogen transmissible via contact, digestive, and respiratory routes, posing severe threats to livestock economies and human health. While its virulence genes critically regulate intracellular survival and replication, the molecular mechanisms underlying pathogenesis remain elusive. This study employed Transposon Directed Insertion-site Sequencing (TraDIS) technology to identify key genes essential for the survival of Brucella melitensis strain 16 M within RAW 264.7 macrophages, resulting in the discovery of 374 anti-phagocytic associated essential genes. Through GO and KEGG analyses, these genes were categorized into eight functional classes, representative genes from each category were selected to generate respective gene-deletion mutants, with bacterial virulence and phenotypes validated via intracellular survival assays, in vitro stress tests, carbon source utilization experiments, and RT-qPCR. All ten constructed mutants exhibited reduced macrophage survival capabilities. Deletion of membrane-related genes cydDC and BME_RS07715 attenuated stress resistance, membrane stability, and immune evasion, knockout of metabolic gene ptsP impaired carbon source utilization and intracellular proliferation, and disruption of regulatory gene BME_RS00125 diminished TCA cycle activity and fatty acid metabolism, compromising energy supply and structural integrity. In this study, some key genes of Brucella surviving in macrophages were verified, and preliminarily elucidated the function of these genes in regulating bacterial virulence, providing a theoretical basis for exploring their mechanism.