Karyotype analysis of Quasipaaspinosa David, 1875 (Anura, Dicroglossidae) with conventional cytogenetic techniques.

The current study analyzed the chromosomal karyotype of Quasipaaspinosa David, 1875 from Hunan Province, China. The karyotype, C-banding, BrdU-banding pattern were characterized using direct preparation of bone-marrow cells and hemocyte cultures. The findings indicated that Q.spinosa was a diploid species (2n = 26) that lacked heteromorphic chromosomes and secondary constrictions. C-banding analysis revealed an abundance of positive signals in the centromere regions, while the BrdU-banding pattern showed three phases in both male and female, occurring consistently and in chronological sequence during S-phase. Notably, there was no asynchronous replication in the late phase. This study enhanced our understanding of the karyotypic structure of Q.spinosa by conventional cytogenetic techniques, thus providing essential scientific insights into the cytogenetics of Q.spinosa.

Myxobolus dumerilii sp. n. (Myxozoa: Myxobolidae) infecting the brain of Chinese longsnout catfish Tachysurus dumerili (Bleeker) in China.

During an investigation of Myxobolus diversity in the Chinese longsnout catfish Tachysurus dumerili (Bleeker), a new species infecting the intracranial epidermis of the host was discovered. Upon opening the cranial cavity, several round whitish plasmodia measuring 0.55-0.80 mm in diameter were observed. Fresh spores (n= 50) were pyriform in the frontal view and fusiform in the sutural view, with a length of 15.4±0.6 (13.9-16.5) µm, width of 9.1±0.4 (8.3-9.8) µm, and thickness of 7.0±0.4 (6.3-7.9) µm. The spores had smooth shell surfaces and transparent membrane sheaths in the posterior. No folds, intercapsular appendix, and caudal appendages were observed. Two equal polar capsules were pyriform and measured 7.5±0.5 (6.7-8.7) µm in length and 3.2±0.3 (2.5-3.6) µm in width. The polar filaments were coiled with five to six turns and perpendicular to the polar capsule length. A BLAST search indicated M. dumerilii sp. n. was closely related to five Myxobolus species (with sequences similarities ranging from 90.54% to 96.52%) found in different organs of yellow catfish Tachysurus fulvidraco (Richardson), rather than the T. dumerili-infecting species M. branchiola Dong and Zhao, 2014 (with 90.5% sequence similarity). Phylogenetic analysis showed that M. dumerilii sp. n. didn't form sister clade with brain-infecting Myxobolus spp, but clustered with M. jianlinensis Gao et Zhao, 2020 and M. voremkhai Akhmerov, 1960 within the Siluriformes-clade with highly supported values, indicating that the host specificity may play a stronger signal than site infections during the evolution of Myxobolus species. Based on the morphological, ecological, and molecular differences observed between the newly discovered species and other available Myxobolus species, M. dumerilii sp. n., is proposed and described in this study.

Protective effects of calcium against cadmium-induced toxicity in juvenile grass carp (Ctenopharyngodon idellus).

Cadmium (Cd) is one of the dominant metal pollutants present in the aquatic environment that affects ion homeostasis, oxidative stress (OS) and immune responses of aquatic organisms. Given the physicochemical similarities between Cd2+ and calcium (Ca2+) ions, their antagonism may facilitate the mitigation of Cd-induced toxicity. To better understand the role of Ca in protecting against Cd-induced toxicity in teleosts, juvenile grass carp were exposed to Cd (measured concentration 3 µg/L) and a gradient of Ca concentrations (measured concentration 1.5 mg/L, 2.5 mg/L, 3.0 mg/L, and 3.5 mg/L in the control (CTL) group, low calcium (LCA) group, medium calcium (MCA) group, and high calcium (HCA) group, respectively) for 30 days. Inductively coupled plasma mass spectrometry (ICP-MS) data analyses showed that simultaneous exposure to Ca impaired the accumulation of Cd in all tested tissues. Besides, Ca addition maintained the plasma ion (Na+, K+, Cl-) homeostasis, alleviated Cd-induced oxidative stress (OS), and regulated the activities and transcriptional levels of ATPase. Furthermore, transcriptional heatmap analysis demonstrated that several indicator genes for OS and calcium signaling pathway were found to be significantly modulated by Ca addition. This work delineates a protective effect of Ca against Cd-induced toxicity in grass carp, providing new insight into the possible solutions to Cd pollution issues in aquaculture industry.

Prediction of Potential Suitable Distribution Areas of Quasipaa spinosa in China Based on MaxEnt Optimization Model.

Quasipaa spinosa is a large cold-water frog unique to China, with great ecological and economic value. In recent years, due to the impact of human activities on the climate, its habitat has been destroyed, resulting in a sharp decline in natural population resources. Based on the existing distribution records of Q. spinosa, this study uses the optimized MaxEnt model and ArcGis 10.2 software to screen out 10 factors such as climate and altitude to predict its future potential distribution area because of climate change. The results show that when the parameters are FC = LQHP and RM = 3, the MaxEnt model is optimal and AUC values are greater than 0.95. The precipitation of the driest month (bio14), temperature seasonality (bio4), elevation (ele), isothermality (bio3), and the minimum temperature of coldest month (bio6) were the main environmental factors affecting the potential range of the Q. spinosa. At present, high-suitability areas are mainly in the Hunan, Fujian, Jiangxi, Chongqing, Guizhou, Anhui, and Sichuan provinces of China. In the future, the potential distribution area of Q. spinosa may gradually extend to the northwest and north. The low-concentration emissions scenario in the future can increase the area of suitable habitat for Q. spinosa and slow down the reduction in the amount of high-suitability areas to a certain extent. In conclusion, the habitat of Q. spinosa is mainly distributed in southern China. Because of global climate change, the high-altitude mountainous areas in southern China with abundant water resources may be the main potential habitat area of Q. spinosa. Predicting the changes in the distribution patterns of Q. spinosa can better help us understand the biogeography of Q. spinosa and develop conservation strategies to minimize the impacts of climate change.

Jirovecia branchilis n. sp. (Microsporidia) from glands of Branchiura sowerbyi (Oligochaeta: Tubificidae) in China.

Jirovecia species primarily infect oligochaetes and are typically characterized by large rod-shaped spores with a tail-like posterior prolongation. Presently, seven Jirovecia spp. are reported worldwide with only one described in China. Here, a new species, Jirovecia branchilis n. sp. was discovered in glands of oligochaetes Branchiura sowerybi Beddard, 1892 in China. Jirovecia branchilis n. sp. elicited the formation of numerous opaque xenomas of 0.12 to 0.20 mm (n = 30) in diameter. Electron microscopic observations demonstrated that the earliest developmental stages observed were uninucleate meronts residing directly with the host cytoplasm. Mature spores were rod-shaped with blunt ends and possessed a collar-like anchoring disk, a manubrium-type polar filament, a bipartite polarplast, and a three-layered spore wall. A tail-like prolongation was distinctly observed in the posterior of spores and measured 13.2-28.6 µm long (n = 30). Jirovecia branchilis n. sp. showed 98.54% sequence similarity with Janacekia tainunus isolated from the fat body of chironomidae larvae Kiefferulus tainanus based on obtained partial SSU rDNA gene sequence, but was significantly different in morphology, host, and infection sites. SSU rDNA-based phylogenetic analysis indicated Jirovecia branchilis n. sp. clustered with Janacekia tainanus within the Jirovecia-Bacillidium-Janacekia clade. In conclusion, a new species within Jirovecia, Jirovecia branchilis n. sp. is erected herein based mainly on its morphological, ecological, and to a lesser degree on its molecular characteristics. The whole relationship between Jirovecia spp., Janacekia spp., and Bacillidium spp. is in need of revision and could potentially be elucidated by using additional makers and sequencing a broader diversity of the already described species.

Gut microbial response to host metabolic phenotypes.

A large number of studies have proved that biological metabolic phenotypes exist objectively and are gradually recognized by humans. Gut microbes affect the host's metabolic phenotype. They directly or indirectly participate in host metabolism, physiology and immunity through changes in population structure, metabolite differences, signal transduction and gene expression. Obtaining comprehensive information and specific identification factors associated with gut microbiota and host metabolic phenotypes has become the focus of research in the field of gut microbes, and it has become possible to find new and effective ways to prevent or treat host metabolic diseases. In the future, precise treatment of gut microbes will become one of the new therapeutic strategies. This article reviews the content of gut microbes and carbohydrate, amino acid, lipid and nucleic acid metabolic phenotypes, including metabolic intermediates, mechanisms of action, latest research findings and treatment strategies, which will help to understand the relationship between gut microbes and host metabolic phenotypes and the current research status.

Two new species of Bacillidium (Microsporidia) from coelomocytes of Branchiura sowerbyi (Oligochaeta: Naididae) in China.

Bacillidium spp. exclusively infect oligochaetes and these microsporidian pathogens are typically characterized by their rod-shaped spores. Seven Bacillidium spp. are presently reported from different organs of oligochaetes. Here, we describe two new Bacillidium species, Bacillidium sinensis n. sp. and Bacillidium branchilis n. sp., from coelomocytes of Branchiura sowerbyi. This is the first report of Bacillidium spp. in oligochaetes from China. Both species of Bacillidium elicit the formations of opaque xenoma-like lesions in coelomocytes of the host. A diplokaryotic nucleus occurs in all life stages of these two new Bacillidium species. Mature spores of B. sinensis are 15.9 ± 0.6 (14.7-17.1) µm long (average ± standard error, range, n = 50) and 2.5 ± 0.1 (2.3-2.7) µm wide in fresh preparations. A new type of exospore (sixteen-layered exospore) is discovered from B. sinensis n. sp. which is distinctly different from B. branchilis n. sp., and other Bacillidium spp. (double-layered exospore) reported previously. These two Bacillidium species are morphologically distinguished from each other and all Bacillidium spp. described previously in terms of hosts, infection sites, spore size, spore wall or polar filament thickness. BLASTn searches indicated that these two new microsporidian parasites are surprisingly most similar to Janacekia tainanus (94.76% for B. sinensis and 90. 2% for B. branchilis) isolated from the fat body of midge larva (Kiefferulus tainanus). Phylogenetic analysis demonstrates that the two novel taxons cluster with J. debaisieuxi, J. tainanus, and Bacillidium sp. within the Jirovecia-Bacillidium-Janacekia clade. Other available 18S rRNA gene sequences for microsporidia that infect oligochaetes include J. sinensis, B. vesiculoformis, Neoflabelliforma aurantiae, and Bacillidium sp., but these do not form a single cluster with B. sinensis and B. branchilis, but are instead dispersed through the clade. Based on the ultrastructural features and molecular characteristics, two new species within the genus Bacillidium, B. sinensis n. sp. and B. branchilis n. sp., are designated.

Naidispora caidianensis n. gen. n. sp. infecting coelomocytes of oligochaete Branchiura sowerbyi (Oligochaeta: Naididae) in China.

A new microsporidian parasite, Naidispora caidianensis n. gen. n. sp. was found in coelomocytes of oligochaete Branchiura sowerbyi Beddard, 1892 from Wuhan city, Hubei Province, China. Opaque, hypertrophied coelomocytes (0.17-0.24 mm in diameter) depicted clinical signs of infection. Electron microscopy revealed a microsporidian with monokaryotic life stages. Rounded uninucleate meronts subsequently transformed into multinucleate merogonial plasmodia with masses of electron-dense projections on their plasma membrane. Electron-dense sporogonial plasmodia separated into uninucleate sporonts through rosette-like budding, and further developed into sporoblasts, enclosed by a sporophorous vesicle. Uninucleate mature spores were pyriform, measured 4.1 ± 0.1 (3.9-4.3) µm × 2.1 ± 0.07 (1.9-2.2) µm (average ± SE, range, n = 50), and contained a mushroom-like anchoring disk, bipartite polaroplast, electron-lucent posterior vacuole, trilaminar spore wall and 12-14 turns of an isofilar polar filament arranged in 2-3 ranks. The 1542 bp sequence from B. soweryi showed the highest similarity (below 80 %) with Nematocenator marisprofundi (accession number: JX463178). SSU rRNA gene-based phylogenetic analysis demonstrated that the novel taxon formed an independent clade from known microsporidian parasites. Based on the ultrastructural features and SSU rRNA gene sequence, we propose the establishment of a novel genus (Naidispora n. gen.) and species (Naidispora caidianensis n. sp.) to contain this parasite.