Morphological Data Research Articles

Morphological and molecular characterization of Eimeria haematopusi n. sp. (Apicomplexa: Eimeriidae) in an Australian Pied Oystercatcher (Haematopus longirostris) (Aves: Charadriiformes).

A novelEimeriaSchneider, 1875 species is described from an Australian pied oystercatcherHaematopus longirostrisVieillot, in Western Australia. The pied oystercatcher was admitted to the Kanyana Wildlife Rehabilitation Centre (KWRC), Perth, Western Australia in a poor body condition, abrasion to its right hock and signs of partial delamination to its lower beak. Investigation into potential medical causes resulted in a faecal sample being collected and screened for gastrointestinal parasites. Unsporulated coccidian oocysts were initially observed in the faeces and identified asEimeriaupon sporulation. The sporulated oocysts (n= 20) are ellipsoidal, 20-21 × 12-13 μm in shape and have thick bi-layered walls which are c.2/3 of the total thickness. Micropyle is present, robust and protruding, and occasionally has a rounded polar body attached to the micropyle. Within the oocyst, a residuum, in addition, two to five polar granules are present. There are four ellipsoidal sporocysts 9-11×5-6 μm with flattened to half-moon shaped Stieda bodies. Sub-Stieda body and para-Stieda body are absent. The sporocysts contain sporocyst residuums composed of a few spherules scattered among the sporozoites. Within the sporozoites, anterior and posterior refractile bodies are present, but the nucleus is indiscernible. To further characterise the novelEimeriaspeciesfromH. longirostris, molecular analysis was conducted at the 18S ribosomal RNA locus, using PCR amplification and cloning. Two cloned sequences from the novelEimeriawere compared with those from otherEimeriaspp. with the highest genetic similarity of 97.6% and 97.2% from Clone 1 and 2, respectively withEimeria reichenowi(AB544308) from a hooded crane (Grus monachaTemminck) in Japan. Both sequences grouped in a clade with theEimeriaspp. isolated from wetland birds, which includeEimeria paludosa(KJ767187) from a dusky moorhen (Gallinula tenebrosaGould) in Western Australia,Eimeria reichenowi(AB544308) and Eimeria gruis (AB544336) both from hooded cranes. Based on the morphological and molecular data, thisEimeriasp. is a new species of coccidian parasite and is named Eimeria haematopusi n. sp. after its hostH. longirostris.

3D intraoral scanning techniques support the effects of crown morphology on dental caries

BackgroundWith the development and utilization of three-dimensional (3D) intraoral scanning (IOS) technology, the morphological characteristics of teeth were quantitatively assessed. In this research, we aimed to explore the prevalence of dental caries in relation to each measurable morphological indicator of the tooth body via 3D intraoral scanning techniques.MethodsA hospital-based single-centre study was conducted at our hospital from Dec. 2021 to Apr. 2023. A total of 53 patients were involved in the study, providing complete morphological data for 79 teeth. Each patient completed an oral hygiene routine questionnaire and underwent examination by an experienced dentist to evaluate caries conditions before undergoing 3D intraoral scanning to obtain a digital dental model. Geomagic Studio 2014 was used to extract oral morphological data from the models. The acquired data were entered, cleaned and edited using Excel 2016 and subsequently exported to SPSS version 25.0 for analysis. Chi-square analysis and logistic regression analyses were employed to test the associations.ResultsAmong the participants, 33 (61.1%) were female, with a mean age of 26.52 ± 10.83 years. Significant associations were found between dental caries and the vertical distance between the distal tip and the gum (OR 14.02; 95% CI 1.80-109.07; P = 0.012), the distal lateral horizontal distance of occlusion (OR 0.40; 95% CI 0.18–0.90; P = 0.026), and the mesial horizontal distance of occlusion (OR 2.20; 95% CI 1.12–4.31; P = 0.021). The Hosmer–Lemeshow test indicated a P value of 0.33.ConclusionsThe vertical distance between the distal tip and the gum, the distal lateral horizontal distance of the occlusion and the mesial horizontal distance of the occlusion were the influencing factors for dental caries (identified as independent risk factors). We hypothesize that these factors may be associated with the physiological curvature of teeth and the role of chewing grooves in plaque formation over time. However, further studies involving larger population samples and more detailed age stratification are still needed.

Marsh-orchids of Canada: long-standing mysteries partially solved

SummaryBetween 1959 and 1988, three populations of purple-flowered terrestrial orchids attributable to Dactylorhiza subgenus Dactylorhiza were discovered in Canada. The populations at Timmins, Ontario, and St John's, Newfoundland were strongly marked on both flowers and leaves, in contrast with the anthocyanin-deficient population at Tilt Cove, Newfoundland. All three populations have since experienced a wide range of taxonomic assignments; debates are also ongoing regarding their origin and most appropriate conservation status. Here, we address these questions by combining detailed in situ morphometric analyses based on 52 characters with allozyme profiles and data from nrITS, 15 plastid microsatellites and seven nuclear microsatellites. The allozyme data alone are sufficient to both confirm allopolyploidy and categorically refute past assignments of these populations to D. incarnata, D. maculata, D. fuchsii, D. majalis or D. purpurella. Several morphometric characters, nuclear microsatellites and nrITS all reliably distinguish each of the three study populations, whereas the two sampled subpopulations from St John's proved near-identical morphologically. In contrast, morphological variation within each of the three populations is strikingly low, particularly in characters other than those influenced by plant vigour. Similarly, compared with 14 European populations, the three Canadian populations proved genetically impoverished (two were near-invariant) and likely experienced recent, extreme genetic bottlenecks during establishment. The three populations differ substantially, both morphologically and molecularly, therefore probably representing independent immigration events. Although clearly attributable to D. praetermissa, all three populations deviate significantly in morphology and DNA data from comparable populations sampled across Europe, preventing identification of their precise geographic origins. Any attempt to determine their mode or origin — through natural long-distance transport, or accidental or deliberate introduction by humans – is challenged to explain why three lineages of a single European Marsh-orchid species, each in different ways atypical of that species, arrived independently in North America whereas no other European dactylorchid species has become established there.

Automated body length and body condition measurements of whales from drone videos for rapid assessment of population health

AbstractMonitoring body length and body condition of individuals helps determine overall population health and assess adaptation to environmental changes. Aerial photogrammetry from drone‐based videos is a valuable method for obtaining body length and body condition measurements of cetaceans. However, the laborious manual processing of drone‐based videos to select frames to measure animals ultimately delays assessment of population health and hinders conservation actions. Here, we apply deep learning methods to expedite the processing of drone‐based videos to improve efficiency of obtaining important morphological measurements of whales. We develop two user‐friendly models to automatically (1) detect and output frames containing whales from drone‐based videos (“DeteX”) and (2) extract body length and body condition measurements from input frames (“XtraX”). We use drone‐based videos of gray whales to compare manual versus automated measurements (n = 86). Our results show automated methods reduced processing times by one‐ninth, while achieving similar accuracy as manual measurements (mean coefficient of variation <5%). We also demonstrate how these methods are adaptable to other species and identify remaining challenges to help further improve automated measurements in the future. Importantly, these tools greatly speed up obtaining key morphological data while maintaining accuracy, which is critical for effectively monitoring population health.

Phenotypic convergence is more frequent and stronger in herbivorous freshwater fishes.

Constraints on phenotypic evolution can lead to patterns of convergent evolution, by limiting the 'pool' of potential phenotypes in the face of endogenous (functional, developmental) or exogenous (competition, predation) selective pressures. Evaluation of convergence depends on integrating ecological and morphological data within a robust, comparative phylogenetic context. The staggering diversity of teleost fishes offers a multitude of lineages adapted for similar ecological roles, and therefore, offers numerous replicated evolutionary experiments for exploring phenotypic convergence. However, our understanding of fish feeding systems has been primarily shaped by marine species, with the monolithic exception of freshwater cichlids. Here we use piranhas and pacus (Serrasalmidae) to explore the evolution of different feeding ecologies and their morphological proxies in Neotropical freshwater environments. Specifically, we explore whether convergence is more widespread among plant-eating fishes, arising from strong constraints on phenotypic evolution in herbivores. Using osteological micro-computed tomographic imaging (μCT), we describe the major axes of morphological variation in pacus and piranhas, regarding their diet and feeding behaviors. Next, we evaluated whether herbivorous niches are less labile than other dietary guilds and whether herbivorous species' phenotypes evolve at a slower evolutionary rate than other taxa. We then assess how convergent herbivorous taxa are, using three different suites of morphological characters (dental, jaw, and abdominal morphometrics). Ecologically, herbivory is not a dead end, exhibiting similar observed transition rates as those between carnivores and omnivores. However, we documented widespread convergence in herbivores and that herbivores have slower rates of phenotypic evolution than carnivores. Most instances of convergence are found in herbivorous taxa, specifically in frugivores and folivores. Moreover, instances of 'complete' convergence, indicated by positive convergence metrics observed in more than one morphometric dataset, were only found in herbivores. Herbivores do appear to evolve under constrained circumstances, but this has not limited their ecological lability.

First Report of Wilt Caused by Fusarium nanum (FIESC 25) on Mungbean (Vigna radiata) in Pakistan.

Mungbean, Vigna radia (L.) R. Wilczek, is ranked 2nd next to chickpea (Cicer arietinum) in total cultivation and production in Pakistan. In August of 2022 and 2023, mungbean plants (cv. PRI Mung-2018) were found wilting in a field at the Ayub Agricultural Research Institute, Faisalabad, Pakistan. Wilted leaves turned yellow, died, but remained attached to the stem. Vascular tissue at the base of the stem showed light to dark brown discoloration. Roots were stunted with purplish brown to black discoloration. Symptomatic mungbean plants were collected from fields at five different locations (20 samples/location). Disease incidence was similar among the five fields, ranging from 5 to 10% at each location depending upon type of germplasm and date of sowing. For fungal isolation and morphological identification, symptomatic stem and root tissues were cut into ~5 mm2 pieces with a sterilized blade. Tissues were surface-sterilized for one min in a 0.5% sodium hypochlorite solution, rinsed twice in sterilized water, air dried on sterilized filter paper, and aseptically placed on potato dextrose agar (PDA) containing 0.5 g/L-1 streptomycin sulphate. Plates were incubated for 3-4 days at 25 ± 2°C with a 12-h photoperiod. Single-spore cultures were used for morphological and molecular analyses. Isolates on PDA grew rapidly and produced abundant white aerial mycelium that turned off-white to beige with age. Macroconidia were hyaline, falcate, typically 3-to-6 septate with a pointed apical cell and a foot-shaped basal cell, measuring 24.5-49.5 x 2.7-4.7 μm (n = 40). Globose to obovate chlamydospores measuring 5.8 ± 0.5 μm (n = 40) were produced singly or in chains and were intercalary or terminal and possessed roughened walls. The morphological data indicated the isolates were members of the genus Fusarium (Leslie and Summerell 2006). To obtain a species-level identification, a portion of translation elongation factor 1-α (TEF1), the largest subunit of RNA polymerase (RPB1), and the second largest subunit of RNA polymerase (RPB2) region were PCR amplified and sequenced using EF1/EF2 (O'Donnell et al. 1998), Fa/G2R (Hofstetter et al. 2007), and 5f2/7cr (Liu et al. 1999) primers, respectively. DNA sequences of these genes were deposited in GenBank under accession numbers MW059021, MW059017 and MW059019, respectively. The partial TEF1, RPB1 and RPB2 sequences were queried against the Fusarium MLST database (https://fusarium.mycobank.org/page/Fusarium_identification), using the polyphasic identification tool. The BLASTn search revealed 99.9% identity of the isolate to F. nanum (Xia et al. 2019), formerly FIESC 25 of the F. incarnatum-equiseti species complex (MRC 2610, NRRL 54143; O'Donnell et al. 2018). To confirm pathogenicity, roots of 3-5 leaf stage mungbean seedlings were soaked in a 106 spores ml-1 conidial suspension of the fungus for 15 min and then planted in 10 cm pots containing sterilized soil. Mock-inoculated plants with sterile water served as a negative control. Twenty pots that were used for each inoculated and control treatment were maintained at 25 ± 2°C, 14:8 h photoperiod, and 80% relative humidity in a growth chamber. After 15 days, leaf yellowing, internal browning from the base of stems and root discoloration was observed in all the inoculated plants. The uninoculated negative control plants remained asymptomatic. Fusarium nanum was re-isolated from artificially inoculated plants and identified by colony growth, conidial characteristics on PDA and molecular analyses (TEF1). To our knowledge, this is the first report of wilt caused by F.nanum on mungbean in Pakistan. In Pakistan, mungbean cultivation in irrigated areas has increased in recent years. It has been introduced frequently in citrus orchards, crop rotation of maize and sesame, intercropping with sugarcane and as green manure. However, citrus, maize, sesame and sugarcane are also hosts of Fusarium spp. Therefore, this information warrants sustainable crop protection and may have an impact on further interaction of F. nanum with other wilt pathogens.

A revised tribal classification of Papaveraceae (poppy family) based on molecular and morphological data

AbstractPapaveraceae, commonly known as the poppy family, contains many species with horticultural and/or pharmaceutical importance and has been the subject of various molecular studies. However, a widely accepted tribal classification for Papaveraceae remains lacking. In this study, we used a sampling covering all 48 of the currently recognized genera of Papaveraceae. Our study is to employ broad morphological data in combination with sequence data from two nuclear and seven plastid DNA regions to reconstruct phylogenetic relationships within the family. Our analyses indicate that Pteridophylloideae is sister to the clade containing Hypecooideae and Fumarioideae, and these three lineages are in turn sister to Papaveroideae. The alternative topologies constraining Pteridophylloideae as sister to the other poppy species, to Papaveroideae or to Hypecooideae, as recovered in previous studies, are all rejected. Fumarioideae contains nine main clades, and Fumarieae is nested in traditional Corydaleae. Within Papaveroideae, three main clades are recovered and Platystemoneae is embedded within traditional Papavereae. Both topologies constraining traditional Corydaleae or Papavereae as monophyletic are significantly rejected. Within this phylogenetic framework, the evolution of selected characters is inferred and diagnostic morphological characters at different taxonomic levels are identified and discussed. Based on morphological and molecular evidence, a new tribal classification of Papaveraceae is presented, in which 4 subfamilies, 14 tribes (including 7 newly described tribes), and 43 genera are recognized.

Human benign Leydig cell tumor − Biochemical evaluation

Recently we reported expressional alterations in 219 genes and their transcripts in Leydig cell tumors but nowadays there is still a lack of full basic biochemical characteristics of these tumors. The discovery of potential biochemical markers for tumor management from early detection, treatments, and control of therapy results may markedly supplement genetic data. Leydig cell micronodules were obtained from patients with azoospermia who were qualified for testicular biopsy. The biochemistry of Leydig cell tumors was analyzed using histological staining and spectrophotometric measurements of total proteins, carbohydrates, lipids, and nucleic acids. In addition, the levels of calcium (Ca2 +), copper (Cu2 +), zinc (Zn2 +), and selenium (Se2 +) ions were measured. When compared to healthy testis we revealed, for the first time, that in the interstitial tissue with Leydig cell tumors, great amounts of proteins, carbohydrates, lipids, and acids were dislocated from the seminiferous tubules. Measurements of organic compounds showed a decrease (P < 0.05) only in the Cu2 + content in Leydig cell tumors which may be related to their altered biochemical structure. This specific result may be promising for designing further approaches to manage this tumor based on combining morphological and molecular data.

New records of amphibians from Ha Tinh Province, Vietnam

Since the establishment of the Vu Quang National Park in 2002 in Ha Tinh Province, central Vietnam, several surveys on the amphibian fauna have been undertaken in this protected area. In 2009, Nguyen et al. provided a list of 23 amphibian species from Vu Quang National Park. In addition, a new species was described in 2021 from the National Park, namely Vietnamophryne vuquangensis. As a result of our field surveys in 2013 and 2023, a total of 29 species of amphibians was recorded from the Vu Quang National Park. Six of them are recorded for the first time from Ha Tinh Province, comprising three species of Megophryidae, one species of Dicroglossidae, one species of Ranidae and one species of Rhacophoridae. In addition, we provide morphological data and ecological notes on the aforementioned species.

A New Species of the Genus Takydromus (Squamata: Lacertidae) from Northeastern Guangxi, China

During our collecting trip to Guangxi in 2016, we collected ten specimens of the genus Takydromus from the suburb of Guilin, northeastern Guangxi, South China, and found that they did not belong to any currently known species. Here, we described this new species, Takydromus guilinensis sp. nov., based on morphological and mitochondrial DNA (CO1 and cyt b) data. This new species is a sister taxon to T. intermedius with a p-distance of 0.070 in CO1 and 0.080 in cyt b. These two p-distances exceed not only the minimum value (0.067) between T. septentrionalis and T. stejnegeri but also the minimum value (0.079) between T. intermedius and T. yunkaiensis. Morphologically, this new species differs from other currently recognized Takydromus species from the same clade, more evidently in the longitudinal rows of dorsal scales, transverse rows of scales at the mid-body and mensural variables. The description of Takydromus guilinensis sp. nov. increases the total number of Takydromus species to 25, of which 16 can be found in China. Takydromus guilinensis sp. nov. is currently known only from Guilin, Guangxi, South China, where it is sympatric with the other four Takydromus species (T. septentrionalis, T. kuehnei, T. sexlineatus and T. intermedius).

Specialized ovipositor sensilla of Cretaceous wasps (Insecta: Hymenoptera) possibly reveal a unique way of host detection.

Insects have evolved complex sensory systems that are important for feeding, defence and reproduction. Parasitoid wasps often spend much time and effort in searching for concealed hosts with the help of specialized sensilla. However, the early evolution of such behaviour and sensilla is poorly known. We describe two fossil female wasps, †Tichostephanus kachinensis sp. nov. and †Tichostephanus longus sp. nov., from mid-Cretaceous Kachin amber. Phylogenetic analyses based on morphological data retrieved †Tichostephanus as deeply nested within Evanioidea and closely related to extant Gasteruptiidae and Evaniidae. Both of these Cretaceous wasps possess features, e.g. coronal tubercles and flexible ovipositor sheaths, that indicate that they might have laid eggs in wood where their larvae possibly parasitized insect larvae. They have a peculiar and unique 'bottle brush' of sensilla close to the apex of their ovipositor sheaths, which has not been observed in any extant parasitoid wasps. These sensilla comprise many regularly arranged plate-shaped setae, attached in relatively large sockets and with rows of longitudinal ridges. Such specialized sensilla perhaps served to enhance the ability to detect hosts inside wood.

An integrative approach highlights the discrepancy in the genetic, phenotypic, and presumptive taxonomic structure of Phoxinus (Actinopterygii, Leuciscidae, Phoxininae) in Bulgaria.

The present drainage network of Bulgaria is the result of a complex Neogene and Quaternary evolution. Karst, which has developed on 23% of the territory, further complicates the hydrological pattern. Fresh waters of Bulgaria drain into the Black Sea and the Aegean Sea basins and can be roughly divided into the Danube (Middle and Lower Danube), non-Danube Black Sea, East Aegean, and West Aegean hydrological regions. Phoxinus, a small leuciscid fish, has a mosaic distribution in all four of these regions, inhabiting small mountainous and semi-mountainous streams. Based on morphology, it was identified as three species, Phoxinus phoxinus in the Danube, Phoxinus strandjae in the non-Danube, and Phoxinus strymonicus in West Aegean region. Later, molecular data revealed Phoxinus csikii and Phoxinus lumaireul in the Middle Danube and P. csikii in the Lower Danube. Phoxinus has been the focus of many studies, showing a high molecular and morphological diversity, which is not entirely consistent with previous morphology-only-based taxonomic concepts. In this study, molecular (a mitochondrial marker and a nuclear marker) and morphological data from both historical and recently sampled collections were analysed to assess the applicability of the integrative approach in Phoxinus. The results showed a significant influence of the complex paleo- and recent hydrology on the currently observed genetic structure of the considered populations and species. Furthermore, the study also demonstrated a strong influence of phenotypic plasticity on the morphological analysis of Phoxinus and the lack of a clear differentiation between P. csikii and P. strandjae. A barcoded specimen was designated as neotype to fix the species named P. strandjae in the current taxonomic concept. Finally, a significant discordance between genetically delimited clades and phenotypic groups did not allow a proper delineation of the species distributed in Bulgaria, demonstrating that more molecular markers are needed for further taxonomic study of the Phoxinus complex.

PSI-21 Optimizing sample size for accurate measurements and prediction equations for intestinal morphology in broiler chickens

Abstract Histological measurements of intestinal morphology, including villi height (VH) and crypt depth (CD), are routinely used to evaluate intestinal health and the nutrient absorptive capacity in monogastric animals. These metrics suffer from significant natural variation, and there is no established standard for the number of unique measurements required to robustly estimate these variables for a given animal. Furthermore, the potential utility of additional variables, including villus width (VW), perimeter (VP), or villus area (VA), has not been explored. Therefore, the objective of the current experiment was to estimate the minimum number of measurements required to control the intrasample variance in each variable and determine if more complex or time-consuming metrics, such as VP and/or VA, can be accurately estimated from linear measurements. A set of 1283 intestinal morphology data, including VH, CD, VW, VP, and VA, measured from 68 intestinal tissues (46 ileum and 22 jejunum) of 4 experiments were used in the current study (Figure 1). Retrospective calculation of the CV for all combinations of N measurements per sample was used to determine the impact of measurement number on the quality of data. Data for the jejunum and ileum were pooled together, and the NLIN procedure of SAS was used to estimate the optimal measurement for intestinal morphology measurements using a quadratic broken line. The VA was estimated using geometric calculation with the assumption that VA equals 0.67 × base width × VH. A paired t-test was conducted to compare measured and calculated VA. The optimal number of measurements to minimize within-sample CV for VH, CD, VW, VP, and VA ranged from 10.6 to 10.8. The VH was highly correlated (P &amp;lt; 0.05) with VP (r = 0.95 and 0.87) and VA (r = 0.77 and 0.63) in the jejunum and ileum, respectively (Table 1 and 2). Additionally, there was a significant difference (P &amp;lt; 0.05) between measured (0.110 µm2) and calculated VA (0.086 µm2). In conclusion, this study shows that measuring 11 villi per sample is required for minimizing variations within the jejunal or ileal tissue in broiler chickens. Also, estimating VA based on idealized geometry was found to be inaccurate, necessitating either direct measurement or more complex prediction equations.

Two new species of Dictyota (Dictyotales, Phaeophyceae), D. dimorphosa sp. nov. and D. recumbens sp. nov., based on morphological and molecular investigations

ABSTRACT The brown algal genus Dictyota (Dictyotales, Phaeophyceae) is widespread, particularly in warm temperate and tropical waters, and plays a vital role in coastal ecosystems. This study reassessed the taxonomy of Dictyota, focusing on specimens from Myanmar and Japan, using a combination of morphological and molecular analyses. Molecular phylogenetic analyses using concatenated DNA sequences of rbcL, psaA, psbA, cox1 and cox3 genes revealed that Dictyota specimens from Myanmar, Japan, and other regions, belong to nine different clades, each representing a different species. Based on morphological and molecular data, seven out of these nine clades are identified as known species, while the remaining two clades are described as new species D. recumbens sp. nov. (=D. sp. A) and D. dimorphosa sp. nov. (=D. sp. B). Dictyota recumbens is characterized by the recumbent thallus, with transverse yellowish-green iridescent lines, and the formation of solitary spores irregularly distributed over the thallus. Dictyota dimorphosa is characterized by two morphological forms: decumbent and erect thalli, with bluish iridescence mainly in the decumbent thallus, and the formation of spores in oval ring shapes distributed over both thallus surfaces. Dictyota recumbens was typically found in tropical regions, while D. dimorphosa appears to be a cool-warm temperate species, distributed from northeastern Hokkaido to southern Kyushu with an extension along South Korea. Our molecular phylogenetic analyses have revealed the existence of several species whose names appeared in multiple clades, suggesting the potential for future increases in species diversity and the presence of cryptic species within certain groups.

Taxonomy and phylogeny of the family Suberitidae (Porifera: Demospongiae) in California

This study presents a comprehensive taxonomic revision of the family Suberitidae (Porifera: Demospongiae) for California, USA. We include the three species previously known from the region, document two additional species previously known from other regions, and formally describe four new species as Pseudosuberites latke sp. nov., Suberites californiana sp. nov., Suberites kumeyaay sp. nov., and Suberites agaricus sp. nov. Multi-locus DNA sequence data is presented for seven of the nine species, and was combined with all publicly available data to produce the most comprehensive global phylogeny for the family to date. By integrating morphological and genetic data, we show that morphological characters may be sufficient for regional species identification but are likely inadequate for global classification into genera that reflect the evolutionary history of the family. We therefore propose that DNA sequencing is a critical component to support future taxonomic revisions.

Taxonomic reassessment of the Herpetoreas xenura (Wall, 1907) (Squamata: Serpentes: Natricidae) from Myanmar with description of a new species

The review of the taxonomic status of Herpetoreas xenura species complex, based on morphological and molecular data, revealed a new species from Myanmar, which we describe as Herpetoreas davidi sp. nov. from the Rakhine Yoma Elephant Wildlife Sanctuary in Rakhine State. It is suggested to be a sister species to Herpetoreas pealii and Herpetoreas xenura sensu stricto, and can be separated from the latter species by a combination of morphological and scalation characters, and by its pattern. Another population from the Htamanthi Wildlife Sanctuary, Sagaing Region, was previously confused with H. xenura. However, as a closer examination of its morphology, shows it to be closer to Hebius khasiensis, we herein tentatively refer to this population as Hebius khasiensis, pending further molecular data confirming its taxonomic status. Consequently, we propose to temporarily remove Herpetoreas xenura from the fauna of Myanmar. Further studies, especially in the northwestern region of Myanmar, such as near border with India in Chin Hills and Naga Hills, are required. An updated key for the species of Herpetoreas is also provided.

Study of defensive behavior of a venomous snake as a new approach to understand snakebite

Snakebites affect millions of people worldwide. The majority of research and management about snakebites focus on venom and antivenom, with less attention given to snake ecology. The fundamental factor in snakebites is the snakes’ defensive biting behavior. Herein we examine the effects of environmental variables (temperature, time of day, and human stimulus) and biological variables (sex and body size) on the biting behavior of a medically significant pit viper species in Brazil, Bothrops jararaca (Viperidae), and associate it with the epidemiology of snakebites. Through experimental simulations of encounters between humans and snakes, we obtained behavioral models applicable to epidemiological situations in the State of São Paulo, Brazil. We found a significant overlap between behavioral, morphological, environmental, and epidemiological data. Variables that increase snakebites in epidemiological data also enhance the tendency of snakes to bite defensively, resulting in snakebites. We propose that snakebite incidents are influenced by environmental and morphological factors, affecting the behavior of snakes and the proportion of incidents. Thus, investigating behavior of snakes related to snakebite incidents is a valuable tool for a better understanding of the epidemiology of these events, helping the prediction and, thus, prevention of snakebites.

Occurrence of Dictyosphaeria (Cladophorales, Chlorophyta) species on the Brazilian coast and oceanic islands, based on morphological and molecular data

Abstract Molecular data related to morphology from the multicellular vesiculated genus Dictyosphaeria are scarce. We provide the first molecular data on Dictyosphaeria species from the southwestern Atlantic Ocean, sampled from the Brazilian northeastern coast, and two oceanic islands (Trindade, southeastern Brazil, and Fernando de Noronha, northeastern Brazil). The samples from Brazil were morphologically identified as Dictyosphaeria versluysii and D. ocellata. Our newly generated sequences of D. versluysii formed a monophyletic clade, while their relationship with D. versluysii from the Indo-Pacific Ocean, where the type locality of the species is located, was unresolved. Additionally, the newly generated sequences of D. ocellata confirm its status as a distinct species but reinforce the difficulties in morphologically separating this species. These results might be indicative of cryptic diversity in D. versluysii and highlight the need for molecular data from other geographic locations to understand the diversity and distribution of Dictyosphaeria species.

First Report of Pseudopestalotiopsis ixorae Causing Leaf Spot of Photinia × fraseri in China.

Photinia × fraseri is a well-known ornamental shrub in southern China. In December 2021, we observed leaf spots that were circular to irregular, gray with dark red margins and violet brown with brownish violet edges on the leaves of Photinia × fraseri shrubs in the scenic area of Shenlongtan (28°46'10″N, 115°42'93″E), Jiangxi Province, China. Almost 15% of the leaves in the 1300 m2 Photinia × fraseri planting area were symptomatic. Thirty symptomatic leaves were randomly collected from different plants, and sectioned into 5-mm2 pieces, which were surface-sterilized using 1% NaOCl for 30 s. After rinsing thrice in sterile distilled water and drying, the pieces were transferred onto potato dextrose agar (PDA) and incubated at 28 ℃ for 5-7 days. A total of sixteen morphologically similar isolates were obtained. After incubation on PDA for 20 days, the fungi had irregular edges, were white to pale brown, and had spare aerial mycelium on the surface with irregularly distributed black, gregarious conidiomata. Conidia were fusoid, subcylindrical, straight to slightly curved, 4-septated, slightly constricted at the septa, and 23 to 36 × 6 to 10 μm (mean: 27.6 × 7.7 μm). The morphological characteristics were consistent with the features of Pseudopestalotiopsis species (Maharachchikumbura et al. 2014). The genomic DNA of two representative isolates (JFRL032 and JFRL033) was extracted for further identification. The internal transcribed spacer (ITS) region, translation elongation factor 1-ɑ (tef1-ɑ) and β-tubulin (tub2) genes were amplified and sequenced using primers ITS5/ITS4, EF1-526F/EF1-1567R, and Bt2A/Bt2B, respectively (Maharachchikumbura et al. 2012). The sequences of the two representative isolates were 100% identical to each other. These nucleotide sequences were deposited in GenBank with accession numbers, ON342794 and ON342795 (ITS); ON375851 and ON375852 (tef1-ɑ); ON375853 and ON375854 (tub2). BLASTn searches of the obtained sequences revealed 99%-100% to ITS (MG816316, 478/478 nucleotides), tef1-ɑ (MG816336, 924/926 nucleotides), tub2 (MG816326, 441/442 nucleotides) sequences of the ex-type strain of Pseudopestalotiopsis ixorae (NTUCC17-001.1). Phylogenetic analysis was conducted using the concatenation of multiple sequences (ITS, tef1-ɑ and tub2) with the Maximum likelihood statistics in PhyloSuite v1.2.2 (Zhang et al.2020). The phylogenetic tree showed the two isolates clustered with P. ixorae in a clade with 100% bootstrap support. The isolates were identified as P. ixorae based on morphological and molecular data. To confirm pathogenicity, eight healthy leaves of 3-year-old Photinia × fraseri were surface sterilized, scratched with a pair of sterilized tweezers, and ten µl of conidial suspension (106 conidia/ml) was sprayed on the injured leaves and the control was sprayed with sterile distilled water. Then, All plants were potted in a climate chamber at 25℃ and 85% relative humidity. After 3 days, leaf spot symptoms similar to those described above were observed on inoculated leaves, while the non-inoculated leaves remained symptomless. The pathogen was reisolated from the inoculated leaves to fulfill Koch's postulates and confirmed as P. ixorae by morphological and molecular analysis. It has been reported that P. ixorae can infect the Ixora plant (Tsai et al., 2018). To the best of our knowledge, this is the first report of P. ixorae causing leaf spot on Photinia × fraseri in China. The study provides valuable information for identifying and controlling the leaf spot on Photinia × fraseri.

Hypothesized life cycle of the snow algae Chlainomonas sp. (Chlamydomonadales, Chlorophyta) from the Cascade Mountains, USA.

Chlainomonas (Chlamydomonadales, Chlorophyta) is one of the four genera of snow algae known to produce annual pink or red blooms in alpine snow. No Chlainomonas species have been successfully cultured in the laboratory, but diverse cell types have been observed from many field-collected samples, from multiple species. The diversity of morphologies suggests these algae have complex life cycles with changes in ploidy. Over 7 years (2017-2023), we observed seasonal blooms dominated by a Chlainomonas species from late spring through the summer months on a snow-on-lake habitat in an alpine basin in the North Cascade Mountains of Washington, USA. The Bagley Lake Chlainomonas is distinct from previously reported species based on morphology and sequence data. We observed a similar collection of cell types observed in other Chlainomonas species, with the addition of swarming biflagellate cells that emerged from sporangia. We present a life cycle hypothesis for this species that links cell morphologies observed in the field to seasonally available habitat. The progression of cell types suggests cells are undergoing both meiosis and fertilization in the life cycle. Since the life cycle is the most fundamental biological feature of an organism, with direct consequences for evolutionary processes, it is critical to understand how snow algal life cycles will influence their responses to changes in their habitat driven by climate warming. For microbial taxa that live in extreme environments and are difficult to culture, temporal field studies, such as we report here, may be key to creating testable hypotheses for life cycles.