The specimens were deposited at the Chengdu Institute of Biology, Chinese Academy of Sciences. The holotype was collected at 27.5065° N, 111.4597° E at approximately 350 metres above sea level. All three animals were found on plant leaves near mountain streams with relatively torrential flow. Animal procedures were conducted under a permit from the Chengdu Institute of Biology’s Animal Care Advisory Committee.

For the molecular analysis, researchers extracted DNA from muscle tissue and sequenced fragments of the mitochondrial 16S rRNA and cytochrome c oxidase subunit I, or COI, genes. They combined these new sequences with available GenBank data for related species, aligned them in BioEdit and analysed the concatenated dataset using maximum likelihood and Bayesian inference.

The concatenated dataset comprised 528 base pairs of 16S and 612 base pairs of COI. A GTR + G + I substitution model was selected using the Bayesian information criterion. Maximum-likelihood support was assessed with 1,000 bootstrap replicates. The Bayesian analysis used four Markov chains for 50 million generations, sampled every 1,000 generations, with the first 25% of sampled trees discarded as burn-in.

The maximum-likelihood and Bayesian trees had nearly consistent topologies, although relationships among many broader lineages remained unresolved. The Xinshao specimens formed a distinct lineage within Leptobrachella, with a Bayesian posterior probability of 1.00 and a maximum-likelihood bootstrap value of 100%. This lineage was sister to a clade containing L. wulingensis and L. yongshunensis.

Uncorrected sequence distances also distinguished the Xinshao population from its congeners. The smallest 16S divergence was 2.1% against L. wulingensis, while the minimum COI distance was 8.1% against L. yongshunensis. The authors reported that these values were equal to or greater than the differentiation observed between some recognized species pairs, including L. bijie and L. jinshaensis, and L. dong and L. dushanensis.

The diagnosis identifies L. xinshaoensis by a combination of characters. Males have a snout–vent length of 27.8–31.4 millimetres. Their toes have rudimentary webbing and narrow lateral fringes; the tympanum is distinct and surrounded by distinct tubercles; and the dorsal surface is shagreened with small, raised tubercles and longitudinal ridges. The ventral surface is creamy white with dark brown spots on the chest and margins, and black spots occur on the flanks. When the hindlimbs are flexed at right angles to the body axis, the heels do not meet.

Measurements and external features separate the new species from its closest relatives. Unlike L. wulingensis, whose heels meet when the hindlimbs are flexed, L. xinshaoensis has a tibia–tarsal articulation reaching between the eye and tympanum rather than the middle of the eye. In L. yongshunensis, the heels overlap, the tibia–tarsal articulation reaches the anterior edge of the eye, and the dorsal skin bears sparse large warts, dense small wart grains and, in some specimens, longitudinal ridges. Comparisons with 83 congeners north of the Isthmus of Kra and 27 species south of it also distinguished the new taxon by body size, glandular anatomy, coloration, toe webbing, limb proportions and skin texture.

The type specimens showed variation in preserved coloration. The holotype had a brown dorsum with an inverted triangular marking between the eyes and a W-shaped marking on the anterior mid-dorsum. Black flank spots occurred among two rows of large warts; the supra-axillary gland was orange; and the upper forelimbs were orange with black transverse bars below. Both paratypes were darker dorsally. One had a whiter abdomen, while the posterior abdominal margin of the other was pale purple. Adult males had subgular vocal sacs and femoral adipose glands, but no nuptial pads or spines.

The specific name xinshaoensis refers to Xinshao County, the type locality. The authors propose the English name “Xinshao Leaf-litter Toad” and the Chinese name “Xin Shao Zhang Tu Chan (新邵掌突蟾).” The species is currently known only from the type locality, and the survey found no females, eggs or tadpoles; late March is therefore described as a speculated breeding season rather than an observed one. Given its restricted known distribution and dependence on specific stream-side habitat, the authors recommend assessing its conservation status in future evaluations and advocate more intensive biodiversity surveys in central Hunan, where other cryptic species may occur.

Subject of Research: Description of Leptobrachella xinshaoensis from Xinshao County, Hunan, China, using integrative morphological and mitochondrial DNA analysis

Article Title: Morphological and molecular evidence support a new species of the genus Leptobrachella ( Anura , Megophryidae ) from central China

Article References: Jiang, L., Liu, J., Huang, Y., Zhang, H., & Wang, B. (2026). Morphological and molecular evidence support a new species of the genus Leptobrachella (Anura, Megophryidae) from central China. Zoosystematics and Evolution, 102(1), 253-269. https://doi.org/10.3897/zse.102.175462

Image Credits: AI Generated

DOI: 10.3897/zse.102.175462

Keywords: Leptobrachella xinshaoensis, new frog species, Hunan China, integrative taxonomy, morphological analysis, mitochondrial DNA, 16S rRNA, COI gene, cryptic species, amphibian diversity, Morphological, molecular

The study assessed Z. puerensis sensu lato, a name previously applied to frogs recorded across much of Yunnan and in northern Vietnam. Researchers conducted field surveys in Yunnan from 2020 to 2026 and collected specimens at several locations. They combined these specimens and newly generated mitochondrial sequences with sequences obtained from GenBank, including samples from elsewhere in Yunnan and northern Vietnam, to examine the taxonomy and distribution of the broadly recorded species.

For the molecular analysis, the researchers extracted total genomic DNA from liver tissue and amplified and sequenced a fragment of the mitochondrial 16S rRNA gene. They aligned the sequences with MAFFT 7, calculated average uncorrected p-distances in MEGA 12.0.10, and constructed phylogenetic trees through both Bayesian inference and maximum-likelihood analyses. The outgroups were Theloderma albopunctatum, Rhacophorus rhodopus and Leptomantis gauni. Species partitions were also evaluated with the Assemble Species by Automatic Partitioning method, in which lower scores indicate better-supported partitions.

Both phylogenetic analyses divided Z. puerensis sensu lato into four clades, although the relationships among those clades were not resolved because node support was low. The first included samples from Pu’er City, the type locality of Z. puerensis, and from Baoshan City, Dali Prefecture, Yuxi City and Honghe Prefecture, all in Yunnan. The other clades contained samples from Sa Pa District in Lao Cai Province, Wenshan Prefecture in Yunnan together with Ha Giang Province in Vietnam, and Lincang City in Yunnan.

Uncorrected genetic distances among the four clades ranged from 2.2% to 2.6%. These distances were greater than those between several species pairs already recognized in the genus, including Z. dugritei and Z. hui at 0.5%, Z. duboisi and Z. meimontis at 1.6%, and Z. lishuiensis and Z. zhoukaiyae at 1.5%. The ASAP analysis supported the Wenshan, Lincang and Sa Pa samples as three species distinct from Z. puerensis sensu stricto. The authors treated the four lineages as separate species based on the combined molecular results, genetic-distance comparisons and available morphological evidence.

Zhangixalus liui has a male snout-vent length of 42.1–46.4 mm and is distinguished by a head wider than long, heels that do not meet when the legs are positioned at right angles to the body, and a tibiotarsal articulation that reaches the centre or anterior margin of the tympanum when the hind limb is stretched forward. Its fifth toe is longer than its third toe, and a nuptial pad is present on the first finger. The dorsal surface is green with more or fewer dark-edged brown spots, while large black patches occur in the axilla and groin region. The holotype and paratypes were collected in Bozhu Town, Wenshan City, at 2,540 m in April 2025.

The species is also known from Ha Giang Province, Vietnam, based on GenBank sequences that differed by 0.5% from the Wenshan specimens. At its type locality, frogs were found in mud holes or grass beside a small pond on grassland surrounded by montane broadleaved forest. Advertisement calls and foamy egg nests were recorded in April, and the authors presumed a breeding period from April to June. The authors also noted that Z. liui can change its dorsal colour from green with brown spots to brown or brownish-black, with corresponding changes in its ventral spotting.

Zhangixalus orlovi has a smaller male snout-vent length of 34.0–41.3 mm. Its tibiotarsal articulation reaches the posterior margin or centre of the tympanum, its third and fifth toes are approximately equal in length, and nuptial pads occur on the first and second fingers. Its dorsal surface may be green with no spots, a few spots or many dark-edged brown spots, while the posterior thigh is black with many white spots. The holotype and paratypes were collected at Daxueshan Township, Yongde County, at 2,940 m in May 2026. This species is currently known only from that type locality, where advertisement calls and foamy egg nests were observed in April and May.

Z. liui and Z. orlovi can be distinguished from one another by body size, toe proportions, the presence of nuptial pads on the first or first and second fingers, and the size and arrangement of black markings on the axilla, groin and posterior thigh. The study remains limited by its reliance on a single mitochondrial gene fragment. The authors said additional gene fragments, particularly nuclear genes, would be needed for more reliable results. They also cautioned that internal and external vocal sacs can be difficult to distinguish from external appearance alone, while the taxonomic status of populations reported from northern Yunnan remains unresolved.

Subject of Research: Taxonomic and genetic assessment of four lineages within Zhangixalus puerensis sensu lato

Article Title: Taxonomic assessment of Zhangixalus puerensis (He, 1999) sensu lato ( Anura , Rhacophoridae ), with description of two cryptic species

Article References: Liu, S., Bu, C., Deng, T., Bai, J., Luo, B., Hou, M., Li, X., Zhang, J., Li, W., Wang, Z., Feng, Y., Wang, J., & Rao, D. (2026). Taxonomic assessment of Zhangixalus puerensis (He, 1999) sensu lato (Anura, Rhacophoridae), with description of two cryptic species. Biodiversity Data Journal, 14, Article e204046. https://doi.org/10.3897/bdj.14.e204046

Image Credits: AI Generated

DOI: 10.3897/BDJ.14.e204046

Keywords: Zhangixalus puerensis, cryptic species, tree frogs, mitochondrial 16S rRNA, phylogenetics, species delimitation, Yunnan, Vietnam, taxonomy, amphibians, Taxonomic, assessment

Boulenophrys , a group of frogs within the subfamily Megophryinae, currently comprises 81 recognised species distributed from eastern and central China across south-eastern Asia to the Sunda Shelf islands, the Philippines, and westwards to Nepal and north-eastern India. Fifty-seven of those species have been described in the last decade alone, and as many as ten new species were published in 2025 by various research groups. The path to the new species began in June 2023, when the survey team collected seven adult males and one adult female from a mountain stream surrounded by evergreen broadleaved forest.

The specimens were euthanised using isoflurane, fixed in 10 percent buffered formalin, and later transferred to 75 percent ethanol for long-term preservation. Muscle tissue for molecular work was stored in 95 percent ethanol at minus 20 degrees Celsius. All voucher specimens were deposited at the Moutai Institute in Guizhou Province, and the survey and handling procedures were approved by the Forestry Bureau of Tianlin County and the Laboratory Animal Ethics Committee of the Moutai Institute.

To determine where the mystery toads sat on the amphibian family tree, the researchers extracted DNA from muscle tissue and amplified two mitochondrial markers: a 525 base pair fragment of the 16S rRNA gene and a 558 base pair fragment of the cytochrome c oxidase subunit I gene, or COI. These were concatenated into a single 1,083 base pair sequence for phylogenetic analysis. The team assembled and aligned sequences using the ClustalW module in BioEdit, selected the best partition scheme and evolutionary model with PARTITIONFINDER, which settled on the GTR plus G plus I model for each partition, and then reconstructed evolutionary relationships using both Maximum Likelihood methods in PhyML and Bayesian Inference in MrBayes.

All Boulenophrys samples formed a well-supported monophyletic clade, and the Tianlin lineage emerged as the sister group to B. anlongensis , a species described from Anlong County in neighbouring Guizhou Province, with maximal Bayesian posterior probability of 1.00 and a bootstrap support of 98 percent. Genetic distances told the same story: within the undescribed taxon, 16S divergence was zero and COI divergence stayed below 0.2 percent, but between the Tianlin toads and B. anlongensis the distance reached 1.4 percent for 16S and 8.2 percent for COI. Those figures are comparable to divergence levels considered valid for species delimitation in congeneric anurans, and they stand in contrast to some closely related species pairs in the genus, such as B. binlingensis , B. yezhongensis and B. binchuanensis , which differ by only 0.3 percent at 16S.

Morphology provided the second line of evidence. The researchers measured seventeen morphometric characters on the eight Tianlin specimens and on five male B. anlongensis , using dial calipers accurate to 0.1 millimetre. Mann-Whitney U tests on the raw measurements showed that B. anlongensis was significantly larger in snout-vent length and in most other traits, with p-values of 0.004 for many characters including head width, tympanum diameter, hind-limb length and tibia width.

Only interorbital space, eye diameter and thigh length failed to reach statistical significance, with thigh length approaching the threshold at p equals 0.051. Because much of this difference could simply reflect overall body size, the team corrected each character for allometry by dividing it by snout-vent length and log-transforming the result before running a principal component analysis. Even after removing the effect of body size, the two species remained clearly separated.

The first two principal components of the size-corrected male data accounted for 53.8 percent of total variance, with the first component, driven largely by interorbital space, hind-limb length and lower arm diameter, explaining 36.3 percent and the second, dominated by head width, explaining 17.5 percent. Specimens of the new species and B. anlongensis clustered apart along the first component axis, confirming that the two taxa differ not only in size but in body proportions as well. The diagnostic features of Boulenophrys tianlinensis are a distinctive combination.

It is a medium-sized frog, with males measuring 34.3 to 39.0 millimetres in snout-vent length and the single known female reaching 47.1 millimetres. Each upper eyelid margin bears one small horn-shaped tubercle. Vomerine ridges are present but vomerine teeth are absent, the tongue is not notched posteriorly, and the tympanum is rounded and clearly visible.

The toes carry only rudimentary webbing and lack lateral fringes, the heels overlap when the thighs are held perpendicular to the body axis, and the tibiotarsal articulation reaches the middle of the eye when the leg is stretched forwards. Males possess a single subgular vocal sac, and breeding males develop dark brown nuptial spines on the bases of the first and second fingers. Compared directly with B. anlongensis , the new species is significantly smaller in nearly every measured trait, lacks the narrow lateral fringes on its toes, and shows a different relative finger length order.

The holotype, an adult male with the catalogue number MT TL20230603004, was collected by Jing Liu on 3 June 2023 at the type locality. In life, the animal displays a dark brown dorsum with an inverted triangular brown speckle between the eyes, X-shaped ridges on the back, four transverse bands on the thighs and shanks, alternating dark and white bars along the lip margins, and a brown-and-white mottled belly. Individual variation among the paratypes is considerable, ranging from darker animals with indistinct striping to lighter ones with prominent brown patches, and the female carries dense black spines on the warty granules of her dorsum and flanks.

The species appears to favour stones in streams and gorges within evergreen broadleaved forest, where it lives alongside at least four other amphibian species, including Boulenophrys jingdongensis , Kurixalus odontotarsus , Amolops sinensis and Leptobrachella ventripunctata . The discovery also carries a broader message about how new species arise in this corner of Asia. Southwest China is a globally recognised biodiversity hotspot, and the authors argue that its karst canyon landscapes, with deep gorges and isolated peaks, act as robust geographic barriers that restrict gene flow, create microrefugia and drive allopatric speciation, a pattern particularly pronounced in groups like Boulenophrys that disperse poorly and occupy narrow ecological niches.

The straight-line distance between the type localities of the new species and B. anlongensis exceeds 100 kilometres, with the Nanpan River and intervening canyons forming effective dispersal barriers. Much remains unknown about the Tianlin Horned Toad: there are no data yet on its population size, breeding ecology or the threats it may face from climate change and human disturbance, and the authors stress that systematic surveys and long-term monitoring are needed to safeguard the species in these fragile karst ecosystems.

Subject of Research: Description of a new horned toad species, Boulenophrys tianlinensis, from western Guangxi, China

Article Title: A new species of the genus Boulenophrys ( Anura , Megophryidae ) from western Guangxi, China

Article References: Liu, J., Cheng, Y., Shen, T., Feng, C., Mu, L., Zhou, J., Wei, G., & Li, S. (2026). A new species of the genus Boulenophrys (Anura, Megophryidae) from western Guangxi, China. Zoosystematics and Evolution, 102(5), 1139-1157. https://doi.org/10.3897/zse.102.193995

Image Credits: AI Generated

DOI: 10.3897/zse.102.193995

Keywords: Boulenophrys tianlinensis, horned toad, new species, Guangxi, China, amphibian taxonomy, megophryid frogs, molecular phylogenetics, karst biodiversity, 16S rRNA, COI gene, cryptic species

The animals were found during fieldwork in a part of the park previously reported to support Acanthosaura lepidogaster. Researchers did not encounter that species at the surveyed high-elevation site. Instead, the collected lizards formed a consistent morphotype distinguished by unusually small nuchal and dorsal crest spines, described as the smallest recorded in the genus. Photographs documented their coloration in life before the animals were euthanized by freezing at minus 10 degrees Celsius for one week, a procedure selected because the authors stated that the species can withstand several days at low temperatures. The specimens were fixed in 10% neutral buffered formalin for one week, rinsed and transferred to 70% ethanol. Liver samples were preserved separately in absolute ethanol for DNA extraction.

A. syrax is a small agamid with a narrow, moderately tall head, a somewhat compressed body and a distinctive arrangement of scales and spines. Its tympanum is small, rounded and scaleless, and the gular pouch is less developed than in many members of the genus. A tiny conical spine sits above the posterior margin of the eye, and a second small spine occurs on the occiput between the tympanum and nuchal crest. The nuchal crest has six or seven short, keeled, compressed and asymmetrical triangular spines. It is separated from the vertebral crest by a narrow diastema of five to eight scales. The dorsal crest itself consists of small, flat, saw-like triangular scales beginning around the shoulders and diminishing toward the base of the tail. The researchers assessed 51 morphometric and meristic characters, including body and limb dimensions, scale counts, spine lengths, tail proportions and the arrangement of head scales.

The type series displays conspicuous differences between males and females. Females reached a maximum snout–vent length of 101.2 millimetres, compared with 69.6 millimetres in males. Among the paratypes, female snout–vent lengths ranged from 75.5 to 101.2 millimetres, while males measured 62.5 to 69.6 millimetres. Female tails were 85.5–106.9 millimetres long and male tails were 72.4–77.6 millimetres long. In both sexes, tail length was relatively short, ranging from 1.06 to 1.16 times snout–vent length. Females lacked the transverse bar found across the dorsal head of the male holotype and had a pale yellowish-brown to orangish-brown body. The male’s body was dark brown with a marbled reticulum and pale greenish spots. Both sexes possessed pale whitish or whitish-yellow markings at the knees and elbows, as well as a black, diamond-shaped nuchal collar.

Other elements of the living coloration also helped distinguish the specimens. The head was dark brown and crossed by creamy-brown transverse bars, with the most prominent bar passing through the orbital region. Male lips and neck were yellowish-green, whereas those of females were whitish-brown. Neither sex had the black eye patch found in several Thai congeners. The underside was creamy, and the male tail was banded in pale brown and dirty yellowish-rusty tones, compared with a yellowish-creamy tail in females. The everted hemipenes of three males measured 5.3–7.2 millimetres in length. Each was symmetrical and spongy, with a budded-rosette shape and a tip width of 3.5–4.3 millimetres; in preserved ethanol, they were creamy yellow.

For the molecular analysis, the researchers extracted DNA from liver tissue and amplified a 698-base-pair fragment of the mitochondrial ND2 gene and a 600-base-pair fragment of COI. They compared the new sequences with sequences representing other Acanthosaura taxa and used Calotes emma as the outgroup for the ND2 analysis. Model selection identified the GTR+I+G substitution model for both datasets. Maximum-likelihood trees were reconstructed in IQ-TREE, with nodal support assessed through 1,000 ultrafast bootstrap replicates. Bayesian analyses in MrBayes used two independent runs, each with four chains, for one million generations; trees were sampled every 1,000 generations, and the first 25% were discarded as burn-in. The investigators defined significant clade support as an ultrafast bootstrap value of at least 95% and a Bayesian posterior probability of at least 0.95.

Both gene trees recovered the eight A. syrax specimens as a monophyletic lineage separated from the other sampled taxa, but the strength of support differed between markers. In the COI analysis, the lineage received 99% maximum-likelihood bootstrap support and a Bayesian posterior probability of 1.00, meeting the study’s thresholds for strong support. The ND2 analysis also produced a monophyletic group and separated it from A. lepidogaster, the closest sampled morphologically similar congener, but its reported support was lower: 76% bootstrap support and a posterior probability of 0.78. The results therefore provided concordant phylogenetic separation, with stronger statistical support from COI than from ND2. The study assessed mitochondrial markers and sampled taxa, rather than examining nuclear genes or every population potentially belonging to the species.

Genetic variation within the new species was minimal. Pairwise differences among the eight specimens ranged from 0 to 0.01% for ND2 and from 0 to 0.34% for COI. In the smaller ND2 comparison with seven other Thai congeners, distances from A. syrax were 0.14–0.21%. Its nearest ND2 comparison was with A. lepidogaster, at 0.14–0.16%. The COI dataset included 14 other congener species, many represented by samples from Vietnam, and showed distances of 14.96–28.13% from A. syrax. The nearest sampled COI comparison was with A. capra, at 14.96–15.44%, while A. lepidogaster differed by 16.64–17.53%. The unusually low divergence in ND2 means the formal recognition of A. syrax depended on the combined evidence of morphology and phylogenetic placement rather than on a single large genetic-distance threshold.

Field observations indicate that A. syrax occurs in a restricted high-elevation habitat within Mae Wong National Park. The researchers encountered it mainly in the upper mountain range above 1,300 metres and did not find individuals below 1,000 metres during their surveys. It was active from early to mid-day, basking on exposed rocks, fallen logs, small shrubs or cement structures. Toward the end of the afternoon, it retreated into the forest to rest beneath logs or rocks or in tree cavities. Its low daytime activity and relative immobility sometimes allowed the researchers to approach and capture specimens by hand without provoking an escape response. At dawn, some individuals were observed feeding on earthworms on the ground.

The species showed marked seasonal variation in activity. It was encountered most frequently, and at high intensity, from April to mid-June, covering the late dry season and transition into the early monsoon. Activity declined as the monsoon progressed and temperatures dropped, and was especially low in November and December, when minimum temperatures reached approximately 10 degrees Celsius. During the rainy season, sightings were concentrated around midday in direct sunlight, suggesting that solar radiation became a limiting factor for activity in wetter, cooler months. The authors interpret the small body size and unusually reduced crest spines as characteristics consistent with a high-altitude lineage, while emphasizing that the known distribution is based on field observations within one national park and has not yet been established across its full potential range.

The discovery shows that the diversity of mountain horned dragons in Thailand has been underestimated and that the highlands of Southeast Asia can retain unrecognized lineages. The authors proposed the English common name “Syrax mountain horned dragon” and the Thai name “King Ka Khaow Naam Lek Syrax.” The specific epithet refers to Syrax, the dragon associated with Queen Rhaenyra Targaryen in the fictional world of A Song of Ice and Fire, reflecting the lizard’s yellowish coloration, diminutive size and reported high clutch size. Because the documented population is associated with fragmented high-elevation forest in Mae Wong National Park, the authors recommend strengthening protection of the park’s primary evergreen forest. They also call for surveys in neighboring mountain ranges to establish the species’ full distribution and determine whether additional cryptic diversity remains undiscovered. The study provides no direct assessment of formal conservation status, but it identifies the known locality as a priority for further fieldwork and habitat protection.

Subject of Research: Morphological and mitochondrial DNA description of a new high-elevation mountain horned dragon from western Thailand

Article Title: Acanthosaura syrax sp. nov., a new species of mountain horned dragon from high-elevation habitats of western Thailand ( Squamata , Agamidae )

Article References: Trivalairat, P., Trivalairat, K., Moungkhaek, N., Caponov, S., & Lorsunyaluck, B. (2026). Acanthosaura syrax sp. nov., a new species of mountain horned dragon from high-elevation habitats of western Thailand (Squamata, Agamidae). ZooKeys, 1287, 47-76. https://doi.org/10.3897/zookeys.1287.186636

Image Credits: AI Generated

DOI: 10.3897/zookeys.1287.186636

Keywords: Acanthosaura syrax, new species, mountain horned dragon, Thailand, Mae Wong National Park, taxonomy, morphology, mitochondrial DNA, reptile biodiversity, high-elevation forests, cryptic species, Acanthosaura

P. barrassoi was described in 2006 from a single male collected at 2,530 metres in the Maiella Mountains of Abruzzo, Italy. That specimen became the holotype. A female originally associated with the species was subsequently found to be a misidentified and unusually small specimen of Colostygia aqueata. A male collected on Monte Terminillo in 2004, about 100 kilometres from the type locality, was barcoded in work reported in 2019 and showed 8.9% sequence divergence from P. incultaria. That isolated genetic result supported recognition of a separate Central Italian taxon but did not establish its wider distribution. The later study brought the type anatomy, mitochondrial barcodes, nuclear genomic data, adult morphology and ecological records together to reassess populations assigned to the P. incultaria–P. barrassoi complex.

Researchers combined external characters with dissections of male and female genitalia, observations of phenology and vertical distribution, and DNA evidence. They measured anatomy with a calibrated microscope eyepiece and examined prepared genitalia and abdomens. For barcoding, they extracted DNA from one or two legs of dried specimens and analyzed a 658-base-pair region of the mitochondrial cytochrome c oxidase I gene, or COI. Sequence divergences were calculated with the Kimura two-parameter model and analyzed using a neighbor-joining algorithm. High-throughput sequencing was also used for older museum material. In parallel, ddRAD sequencing generated genome-wide single-nucleotide polymorphisms, or SNPs, for maximum-likelihood phylogenetic analysis and population-clustering tests. The authors used these independent evidence streams together because the two moths have no constant and reliable external wing feature that permits separation in the field.

The mitochondrial results divided the tested specimens into two clearly separated groups. The Monte Terminillo specimen produced a 658-base-pair sequence assigned to BOLD bin ACJ5976. Six specimens from Austria, southwestern Bavaria and northern Italy shared that bin and were only 1.1% distant from the Central Italian sequence. Their nearest species was P. incultaria, represented in the comparison by 19 specimens from Germany, Austria and northern Italy in bin AAF5044, with a reported barcode distance of 8.5%. Two specimens attributed to P. incultaria from the vicinity of Gastein in Salzburg were also successfully sequenced using high-throughput methods and fell unambiguously into the P. incultaria bin. Thus, the barcode evidence separated the taxa while also connecting Alpine populations with the Central Italian reference material.

Male genital anatomy strongly supported the DNA assignments. The newly attributed Alpine and Pyrenean populations closely matched the Italian holotype of P. barrassoi. In this species, the valvae are comparatively narrow, with their dorsal and ventral margins approximately parallel, and the costa narrows toward a finely tapered apex. Its aedeagus bears five to nine small cornuti measuring approximately 0.1 to 0.25 millimetres, none expanded at the base. P. incultaria has a much broader valval costa that dilates toward its apex and terminates in a small, beak-shaped sclerotized process bearing a conspicuous tuft of setae. Its aedeagus has five to seven stouter cornuti, with the distalmost generally the largest, curved and dilated at the base. The diagnostic contrasts were assessed in 7 male P. barrassoi and 23 male P. incultaria dissections.

Female genitalia supplied additional but subtler distinctions. In the six P. barrassoi females examined, the apophyses are fine and tapered, while the elongated colliculum is almost parallel-sided for most of its length. Its reported length was 0.37–0.5 millimetres in most specimens, with one measuring 0.6 millimetres. The corpus bursae is small and often globular, and the scobinate signum is small and round, with weak lateral spinules measuring 0.10–0.17 millimetres in diameter. Female P. incultaria has a generally longer colliculum of 0.6–0.7 millimetres and a larger, more elongate and often triangular signum of 0.13–0.3 millimetres, accompanied by stout lateral spinules. The sexes therefore differed in several reproductive structures, although the strongest visible separation was in the male genitalia.

Genome-wide data independently recovered the same two lineages. The ddRAD workflow generated an average of 1.54 million reads per individual, of which 84.8% remained after stringent quality filtering. A de novo assembly recovered 1,042 putative orthologues shared by more than three samples, covering 193,994 base pairs in total. This dataset contained 1,678 SNPs, including 200 parsimony-informative sites. A reference assembly mapped against the genome of the distantly related moth Operophtera brumata yielded a smaller final dataset of 50 retained loci across more than three individuals. Maximum-likelihood trees constructed independently from the de novo and reference datasets produced congruent arrangements, with the P. incultaria and P. barrassoi lineages receiving 100% bootstrap support. Population-clustering analysis likewise identified two discrete genetic groups corresponding to the two species. Only one of the eight individuals tested carried the bacterial parasite Wolbachia.

The revision also addressed the uncertain name-bearing material of P. incultaria. Its original description was based on two males and one female from the Alps. Two collectors were mentioned, with Gastein in Salzburg associated with the material, but the type specimens were presumed destroyed and could not be traced in museums in London or Berlin. Because P. incultaria cannot be separated externally from P. barrassoi and both may occur around Gastein, the authors designated a neotype to preserve nomenclatural stability. The chosen male was collected at Bad Hofgastein, Schlossalm, at 2,000 metres on 6 August 1975. High-throughput sequencing recovered a 658-base-pair fragment that clustered unambiguously with P. incultaria, fixing the name to a documented, genetically identified individual.

Ecological and seasonal records suggest reasons why P. barrassoi may have escaped attention. It is a montane, univoltine species recorded from approximately 700 metres in Bavarian and Austrian valleys to 2,500 metres in the Maiella Mountains. Other sites span about 1,000–1,300 metres in the Bavarian and Austrian Alps, 1,500–2,100 metres in the French Alps and Pyrenees, 1,800 metres on Monte Terminillo and 2,450 metres on the Adamello. Its central Italian specimens were recorded in mid- and late July, and its Pyrenean and French Alpine flight period extends from early July to early August. Alpine records, however, range from mid-April to late May, with one eastern Tyrolean specimen from mid-June and northern Italian specimens from late June. Where the two species coexist, P. barrassoi generally flies earlier than P. incultaria, whose Alpine populations usually fly from early June to early August. The larval stages and host plants of P. barrassoi remain unknown, although its sister species feeds on Primulaceae in early larval stages and on Saxifragaceae later, and representatives of both plant families occur at P. barrassoi collection sites.

On the present evidence, P. barrassoi is confirmed in the central Apennines, the Pyrenees, the French Alps, the Adamello area of northern Italy, eastern and northern Tyrol in Austria, and southernmost Bavaria in Germany. These confirmed records consist of distinct subareas, and the authors considered incomplete recording a more likely explanation than a genuinely fragmented distribution. They predicted that additional populations could remain in the northern Apennines and the western Alps, including Switzerland, Piedmont and Valle d’Aosta, and noted that the species’ early flight period and close resemblance to P. incultaria could make it easy to overlook. That wider range remains a prediction rather than a documented finding. Within the studied material, however, the combined barcode, nuclear-genomic, anatomical and ecological evidence establishes P. barrassoi as a distinct species occurring alongside P. incultaria across previously unrecognized parts of Central and Western Europe.

Subject of Research: Taxonomic identification and European distribution of the geometrid moth Perizoma barrassoi

Article Title: DNA barcoding and genomics reveal Perizoma barrassoi Zahm, Cieslak & Hausmann, 2006 as new for the fauna of Central Europe ( Lepidoptera , Geometridae , Larentiinae )

Article References: Hausmann, A., Huemer, P., Lee, K. M., & Mutanen, M. (2021). DNA barcoding and genomics reveal Perizoma barrassoi Zahm, Cieslak & Hausmann, 2006 as new for the fauna of Central Europe (Lepidoptera, Geometridae, Larentiinae). Nota Lepidopterologica, 44, 17-28. https://doi.org/10.3897/nl.44.58871

Image Credits: AI Generated

DOI: 10.3897/nl.44.58871

Keywords: Perizoma barrassoi, DNA barcoding, ddRAD sequencing, genome-wide SNPs, cryptic species, Geometridae, Lepidoptera, moth taxonomy, genital morphology, European fauna, barcoding, genomics

The findings show that closely similar shrimp can conceal substantial species diversity, while also highlighting the vulnerability of the island’s isolated freshwater habitats. The research focused on a nine-member complex centered on Caridina clinata, a small freshwater shrimp originally described in 1999 from Cuc Phuong National Park in northern Vietnam. Its close relative C. haivanensis was described a decade later from the Hai Van Mountain Pass in central Vietnam.

Phylogenetic analyses based on the mitochondrial cytochrome c oxidase subunit I gene, or COI, and the 16S ribosomal RNA gene resolved the complex into nine distinct, well-supported clades. C. clinata was recorded from both Vietnam and China, C. haivanensis remained restricted to Vietnam, and the seven newly described members of the complex were endemic to Hainan. Hainan is a continental island of 33,900 square kilometres in the South China Sea and the second-largest island in China.

Its high central terrain and lower surrounding areas produce a radial river system, including the Nandu, Changhua and Wanquan rivers, which together drain more than 3,000 square kilometres. The island’s varied geology, climate and habitats spanning marine, brackish and freshwater environments have contributed to high endemism. The newly described shrimp are locally distributed: Caridina fontinalis was found at a spring outlet near Haikou, while C. wenchangensis, C. qiongzhongensis, C. huangi, C. hepingensis, C. tunchengensis and C. brevilineata were each recovered from streams associated with particular towns in Wenchang, Qiongzhong, Tunchang and Qionghai.

Fieldwork was conducted from 2023 to 2025 at 14 sites, comprising 12 locations on Hainan and two in Vietnam representing material of C. clinata and C. haivanensis from their type regions. Researchers used fine-mesh dip nets and baited traps, recorded GPS coordinates and photographed habitats and live animals, and measured site conditions including substrate, water current, pH, dissolved oxygen and ammonia nitrogen. Specimens were preserved in ethanol and examined with a stereomicroscope.

For molecular analysis, DNA was extracted from abdominal muscle, the COI and 16S loci were amplified and sequenced, and the aligned sequences were trimmed to 490 and 473 base pairs, respectively. A concatenated maximum-likelihood analysis used the GTR+G+I model, with branch support evaluated through 1,000 bootstrap replicates. The eight new species were named Caridina wenchangensis, C. qiongzhongensis, C. huangi, C. hepingensis, C. fontinalis, C. tunchengensis, C. brevilineata and C. caii.

Their recognition rested on combined evidence rather than a single diagnostic character. Relevant traits included the length and tooth armature of the rostrum, proportions of the walking legs, form of the male reproductive appendages, structure of the telson, uropodal spines and egg dimensions. For example, C. fontinalis, described from cool water with stable flow, has a shorter rostrum and fewer teeth than C. lanceifrons, with which it occurs at the same sampling site.

C. brevilineata, whose name refers to short body stripes, differs from C. clinata in its leaf-shaped male first pleopod endopod, reduced rostral armature and the absence of a finger-like projection on the first maxilliped palp. The Wuzhishan population supplied the clearest instance of a revised identification. It had been assigned to C. clinata by Cai in 2014 because of its resemblance to C. lanceifrons, but new specimens from the locality and topotypic C. clinata from Vietnam showed that it was distinct in both morphology and genetics.

The Wuzhishan shrimp differed from C. clinata by 25.4–29.3% in COI sequence divergence and 10.8–12.1% in 16S divergence. It also possessed a pointed, subtriangular male first pleopod endopod rather than the rounded form found in C. clinata, a relatively broad second walking-leg carpus, fewer spines on the fifth leg and smaller eggs. The researchers named the species C. caii in honour of carcinologist Cai Y.

X., who discovered the species and made substantial contributions to carcinology. Genetic evidence also supported retaining the Ledong population of Hainan within C. clinata. Shrimp from Ledong differed from Vietnamese populations by only 0.6–1.2% in COI sequences and 0.7–1.6% in 16S, and the two populations occupied the same well-supported phylogenetic clade.

Although some morphological measurements varied, the male first pleopod and second pleopod appendages were closely alike. By contrast, C. wenchangensis was genetically closest to C. tunchengensis and C. clinata but remained a separate supported lineage. C. huangi and C. hepingensis, collected from unconnected streams in the same county, were likewise distinguishable in both morphology and genetic distance.

Members of the C. clinata complex occupy the middle and upper reaches of mountain streams. They shelter beneath leaf litter, among marginal vegetation or on muddy, sandy, gravelly and rocky substrates, generally in slow- to moderately flowing water. The complex produces relatively few large eggs, a reproductive pattern associated with direct larval development into benthic juveniles and an exclusively freshwater life cycle.

Field observations recorded dense populations in several streams, including the Ledong, Wuzhishan, Qiongzhong, Heping and Tuncheng localities, but lower densities at the Wenchang, Haikou and Qionghai sites. The study sampled only a limited set of streams, so the geographic range and population trends of the newly described taxa remain incompletely characterized. The survey also documented pressures on the habitats of these isolated shrimp.

Rubber and areca-nut plantations surround many streams, and intensive irrigation has reduced flow in some locations. Temporary in-stream diversions installed during droughts have left sections of streams completely desiccated, while pipelines have been used to extract water for domestic use. Hydropower and irrigation infrastructure further fragments river corridors, and agricultural chemicals threaten water quality.

Their specific effects on native shrimp remain undocumented, although predation, resource competition and disease transmission are recognized pathways through which invasive species can reduce native biodiversity. Following the International Union for Conservation of Nature’s Red List Categories and Criteria, the researchers classified all eight newly described species as Data Deficient because they are known only from their type localities and broader distribution data are unavailable. C. clinata retained the Data Deficient assessment assigned in an earlier evaluation.

These classifications identify major gaps in knowledge rather than low or high extinction risk. The study supports regular monitoring of wild populations and public-awareness efforts emphasizing the ecological importance of freshwater shrimp, while its broader sampling and genetic records provide a baseline for future surveys of Hainan’s stream biodiversity and for reassessing the conservation status of the newly recognized species.

Subject of Research: Taxonomic and phylogenetic study of the Caridina clinata species complex in Hainan and Vietnam

Article Title: Integrative taxonomy reveals the Caridina clinata Cai, Nguyen & Ng, 1999 species complex ( Crustacea , Decapoda , Atyidae ), with descriptions of eight new species from Hainan Island, China

Article References: Hou, J., Zhang, J., Chen, B., Zhang, Y., Chen, W., & Guo, Z. (2025). Integrative taxonomy reveals the Caridina clinata Cai, Nguyen & Ng, 1999 species complex (Crustacea, Decapoda, Atyidae), with descriptions of eight new species from Hainan Island, China. Zoosystematics and Evolution, 101(4), 2295-2336. https://doi.org/10.3897/zse.101.172207

Image Credits: AI Generated

DOI: 10.3897/zse.101.172207

Keywords: Caridina shrimp, Hainan Island, integrative taxonomy, cryptic species, freshwater biodiversity, molecular phylogeny, COI sequencing, 16S rRNA, species delimitation, conservation assessment, Integrative, taxonomy

The more northern and broadly distributed G. pennsylvanicus occurs throughout much of the United States, whereas G. firmus is concentrated along the eastern coast and extends into Texas. The species, whose common ancestry dates to roughly 200,000 years ago, meet along the eastern front of the Appalachian Mountains and form a hybrid zone. They differ in several respects: G. firmus is often associated with sandy habitats, tends to be lighter in color, has longer ovipositors and is generally larger.

G. firmus females mated to G. pennsylvanicus males lay few eggs that do not hatch, an asymmetry consistent with the direction in which mitochondrial DNA moves between the species. These differences had previously been characterized in only a handful of locations, particularly Connecticut. That focus left open whether the same traits consistently identified the species across their full ranges.

The distinction matters because body size can vary with climate and growing-season length, while ovipositor length may be associated with local soil conditions. A trait that appears diagnostic in one hybrid zone could therefore reflect adaptation or population history rather than a stable species difference. To address that problem, the researchers assembled published and unpublished measurements collected with consistent methods, supplemented them with molecular markers for a subset of crickets and compared patterns across broad geographic regions and within the Connecticut hybrid zone.

Among allopatric populations, the two species differed significantly in male and female body size, absolute ovipositor length and ovipositor length relative to body size. The distributions nevertheless overlapped. Ovipositor length provided the clearest separation and remained informative after being scaled relative to pronotum width.

In a small subset of males for which wing-cover color was measured by spectrophotometry, color alone misclassified 3% of individuals as the wrong species: one of 24 G. pennsylvanicus males and none of seven G. firmus males. Body size alone misclassified 23% of 358 males, with 56 of 268 G. pennsylvanicus and 27 of 90 G. firmus assigned incorrectly. For females, body size and relative ovipositor length together produced a 12% misclassification rate, with 15 of 189 G. pennsylvanicus and 17 of 90 G. firmus misclassified.

These results identify useful diagnostic combinations, but they do not show that any one trait cleanly separates every individual. Crickets from or near the hybrid zone overlapped extensively with the allopatric samples, yet the sympatric group was intermediate for several female traits. Its females differed from both allopatric species in body size, absolute ovipositor length and relative ovipositor length.

Male body size in sympatric populations was not significantly different from that of allopatric G. firmus, although it was greater than in G. pennsylvanicus. The findings indicate that populations in contact do not simply reproduce the morphology of either pure species. They also show why a broad geographic survey is necessary: measurements from a single focal area may capture a regional form rather than the full extent of species divergence.

Variation within each species was substantial. G. pennsylvanicus from the southern, southwestern and central United States were larger than populations in the northwest and west-north-central regions. G. firmus from the south were also generally larger than those from the Northeast, although the southern samples were not significantly different from either northeastern or southeastern populations.

Ovipositor length varied geographically within both species. Eastern G. pennsylvanicus had the shortest ovipositors and central populations had the longest, though the central sample was limited. Within G. firmus, relative ovipositor length was shortest in the south, intermediate in the Southeast and longest in the Northeast.

Southeastern females had nearly the same absolute ovipositor length as northeastern females but were larger, showing why absolute and body-size-adjusted measurements can tell different geographic stories. Southern G. firmus populations also differed sharply from one another. Florida animals were much larger than northeastern G. firmus but had only slightly longer ovipositors, producing shorter ovipositors relative to body size.

Texas G. firmus did not differ from northeastern G. firmus in overall size but had still shorter relative ovipositors. The magnitude of these differences was similar to that separating G. firmus from the closely related, recently described G. thinos. Earlier genetic analyses had also placed Texas and Florida G. firmus-like populations in distinct evolutionary clusters.

The combined morphological and genetic evidence supports the proposition that Texas G. firmus may be an undescribed cryptic species, but the study notes that a phylogeny including northeastern, Texas and Florida populations, along with tests of reproductive compatibility, is still needed to resolve their relationships. Genetic data from the hybrid zone connected morphology to ancestry. Among crickets with a mixture of nuclear DNA from the two species, highly admixed individuals generally had intermediate phenotypes.

The nuclear hybrid index, which estimates the proportion of alleles inherited from G. firmus, explained 47% of the variation in relative ovipositor length. Its association was also strong with female body size, male tegmina color and male body size, explaining 28%, 31% and 19% of their respective variation, respectively. The analysis also detected asymmetric mitochondrial introgression: crickets with predominantly G. firmus nuclear ancestry frequently carried G. pennsylvanicus mitochondrial DNA.

Environmental models identified associations but did not reveal a single mechanism accounting for morphology across the entire distribution. Across the species’ ranges, the best-supported model for ovipositor length included latitude, elevation, average soil clay content and minimum and maximum temperatures; the selected model for pronotum width included latitude, longitude, soil sand content and minimum temperature. Higher clay content and higher minimum and maximum temperatures were associated with longer ovipositors, while higher minimum temperatures were associated with larger body size.

Within Connecticut, minimum and maximum temperatures and soil organic matter were retained in the best-supported model for ovipositor length. However, the expected simple relationship between sandy soil and long ovipositors did not hold. Both species use their ovipositors to place eggs in soil, but ovipositor length does not necessarily dictate laying depth, and established soil preferences and egg-viability patterns are geographically localized.

The authors therefore emphasize that habitat associations are variable and require further investigation. Body size also varied in ways compatible with climatic and developmental constraints. Northern populations tended to be smaller, matching the pattern expected where a shorter growing season leaves less time for growth.

G. firmus additionally shows regional variation in seasonal emergence: in Virginia it emerges earlier than G. pennsylvanicus, creating temporal isolation, whereas the two species emerge simultaneously in Connecticut. Florida G. firmus may produce multiple generations annually yet are considerably larger than northern populations, leaving open whether its life cycle differs discontinuously from that of other G. firmus. The geographic study therefore establishes a need to test species boundaries across many populations and environments, but it does not demonstrate that one model of body-size evolution, habitat association or hybrid-zone structure can explain every population.

Its clearest conclusion is more limited and more useful: traits that distinguish two species in one region may shift elsewhere, so apparent divergence must be assessed with range-wide sampling, genetic evidence and explicit consideration of local conditions.

Subject of Research: Geographic variation in morphology, ancestry, and environmental associations in two hybridizing field cricket species

Article Title: Geographic variation in phenotypic divergence between two hybridizing field cricket species

Article References: Byerly, A. R., Jenck, C., Goetz, A. R. B., Weissman, D. B., Gray, D. A., Ross, C. L., Maroja, L. S., & Larson, E. L. (2023). Geographic variation in phenotypic divergence between two hybridizing field cricket species. Journal of Orthoptera Research, 32(2), 189-200. https://doi.org/10.3897/jor.32.90713

Image Credits: AI Generated

DOI: 10.3897/jor.32.90713

Keywords: field crickets, Gryllus firmus, Gryllus pennsylvanicus, phenotypic divergence, hybrid zones, speciation, cryptic species, local adaptation, ovipositor length, mitochondrial introgression, Geographic, variation

The study identifies P. (M.) haennii as a cryptic species whose close relatives cannot be separated reliably from external appearance alone. The investigation formed part of an entomological program conducted by the Natural History Museum of Canton Ticino in Lugano to document forest insect diversity around Locarno. Investigators surveyed three stations in a Querco-castagnetum forest over a full year, from July 2015 to July 2016, and used different kinds of traps.

Three slightly different attractant traps—transparent traps containing wine or beer and a yellow trap containing wine—captured 99.4% of the Periscelididae collected. A total of 305 males of the new species were trapped. The material was initially preserved in ethanol, after which 53 specimens designated as the type series were dried and mounted on pins for examination.

Under the most recent identification key available to the researchers at the time, many of the captured flies were treated as P. (M.) annulata, a species described by Fallén in 1813. However, the male terminalia of the specimens did not match the diagnostic features of that species. This discrepancy prompted taxonomic verification with international expertise from the late László Papp of Budapest and Miguel Carles-Tolrá of Barcelona.

To determine whether the unusual genital structure represented a distinct species, the team examined the male terminalia of all three European members of Myodris: P. (M.) haennii, P. (M.) annulata, and P. (M.) piricercus, described by Carles-Tolrá and Verdugo Páez in 2009. The male terminalia of P. (M.) annulata were redescribed and illustrated as part of that comparison. The holotype of the new species measures 1.31 mm in total body length and 1.01 mm in wing length.

Its general coloration is brown and grey, with dull, microtomentose surfaces and yellow to yellow-ochre areas on the head, mouthparts, and legs. The microtomentum of the scutum and scutellum is darker and browner than in P. (M.) annulata, so the brown acrostichal and dorsocentral stripes have less contrast against the background. The fly’s legs are yellow and brown, while small silvery-white spots occur along the lateral margins of abdominal tergites 3–6.

Although some of these external traits differ from P. (M.) annulata, the decisive features are found in the male genitalia, particularly the cerci, surstylus, postgonite, and ejacapodeme. In the new species, the cerci are longer and more slender than those of P. (M.) annulata and longer than the anal opening of the epandrium. Each cercus bears two short, robust anteroapical spines that arise close together, while its subapical posterior setae are scarcely longer than adjacent posterior setae.

The surstylus narrows more gradually and remains relatively straight distally. The postgonite is more elongate, with a slender distal portion that is longer and slightly bent. The ejacapodeme is also proportionally more elongate, measuring about 5.5 times as long as its maximum width, compared with about 4.5 times that length in P. (M.) annulata.

These distinctions form the basis of the authors’ identification key for European male Myodris flies. Molecular analysis supplied an additional line of evidence. The researchers amplified and sequenced the COI barcoding region from one specimen each of P. (M.) haennii, P. (M.) annulata, and P. (M.) piricercus, together with Periscelis (s. str.) winnertzii for comparison.

All analyzed flies came from Losone in Ticino and had been preserved in 80% ethanol. The COI sequences were 658 base pairs long, and genetic distances were calculated with the Kimura two-parameter model. P. (M.) haennii differed from P. (M.) annulata by 7.58% and from P. (M.) piricercus by 9.12%.

The distance between P. (M.) annulata and P. (M.) piricercus was 6.88%. Because only one specimen per species was barcoded, the analysis could not measure intraspecific variation. Long series of all three European Myodris species from the same locality and habitat also revealed previously unrecognized variation in thoracic micropubescence, color pattern, and clouding of the wing membrane and veins.

The wing clouding was particularly variable in P. (M.) annulata. As a result, the researchers concluded that the three species cannot be recognized safely using only external features and that identification should always be verified by examining the male genitalia. Females consequently cannot be assigned unambiguously to species from morphology at present, although the study reports that molecular barcoding can identify them.

The new key is therefore restricted to males and uses genital characters rather than the variable external coloration and wing pattern seen in the field material. Within the key, P. (M.) piricercus is separated from the other two species by its sparsely setose pregenital sternum, short pyriform cercus with an expanded proximal half, short and broadly based surstylus, and short, broad postgonite with a robust apex. P. (M.) haennii and P. (M.) annulata both have more densely setose pregenital sternites, slender elongate cerci, long and slender surstyli, and long, distally slender postgonites.

The new species differs from P. (M.) annulata in the relative length of its cercus, the close spacing of the two anteroapical cercus spines, the reduced prominence of the subapical posterior setae, the straighter distal surstylus, and the longer, slightly bent distal portion of the postgonite. These are the specific observations used to distinguish the two species under magnification. The type locality is the Forest Reserve of Parco Collina di Maia in Losone, at 46°09′53″ N, 8°44′54″ E and an elevation of 419 metres.

The holotype is a pinned male collected between 21 and 28 July 2015 in a chestnut-and-oak forest and is deposited in the Natural History Museum of Canton Ticino. Paratypes are held in Lugano and at the Silesian Museum in Opava, Czech Republic. Additional material was collected in 2016 and 2017.

At present, P. (M.) haennii is known only from southern Switzerland. The authors suggest that it may also occur elsewhere in Europe, particularly in more southern areas, because earlier records of P. (M.) annulata may have included the new species, but no confirmed distribution outside Ticino is reported. The biology of P. (M.) haennii remains unknown.

The authors consider it plausible that the species has habits similar to P. (M.) annulata, which has been associated with sap runs on wounded trees, especially oaks and elms, often in their crowns, and whose larvae reportedly develop in fermenting tree sap. That comparison is explicitly provisional and does not document the biology of the new species. What is known directly is that 305 specimens were collected in wine- and beer-baited traps in a lowland forest dominated by oaks and chestnuts.

The same traps also captured both other European Myodris species and two species of Periscelis in the strict sense. The authors describe P. (M.) haennii as another cryptic member of the Periscelididae and suggest that additional unnamed species may be found, particularly in less thoroughly explored eastern and southeastern parts of Europe, when long-term fieldwork and specialist taxonomic examination are available.

Subject of Research: Taxonomic identification of a new cryptic Periscelididae fly species using male genital morphology and COI DNA barcoding

Article Title: Periscelis ( Myodris ) haennii sp. nov., a new species of Periscelididae ( Diptera ) from Ticino, Switzerland, with a new key to European species of the subgenus

Article References: Pollini Paltrinieri, L., & Roháček, J. (2022). Periscelis (Myodris) haennii sp. nov., a new species of Periscelididae (Diptera) from Ticino, Switzerland, with a new key to European species of the subgenus. Alpine Entomology, 6, 39-49. https://doi.org/10.3897/alpento.6.85391

Image Credits: AI Generated

DOI: 10.3897/alpento.6.85391

Keywords: new species, Periscelis haennii, Periscelididae, cryptic species, DNA barcoding, male genitalia, Myodris, Ticino, forest biodiversity, taxonomy, Periscelis, haennii

The round-winged katydids belong to the rotundifolia species group of Amblycorypha, whose members are flightless, medium-sized and difficult to distinguish by external appearance alone. They live in longleaf pine and turkey oak sandhills across the Southeast and typically move only about 10–15 metres per day. Earlier research had identified song differences among populations assigned to A. bartrami, including anomalous calls in South Carolina and North Carolina, but the broader investigation combined recordings, standardized measurements of preserved specimens and DNA sequences to assess whether those differences corresponded to separate evolutionary lineages.

Most fieldwork was conducted at night. Researchers located males by listening for their calls and searching vegetation with headlights, while some females and males were collected during the day with sweep nets. Calling songs of free-ranging or caged males were recorded using shotgun microphones and digital recorders at sampling rates of 44 or 48 kilohertz.

For laboratory recordings, microphones were placed 0.5 metres from the males, with foam placed between the animals and the microphones to reduce echoes and background noise. Because katydid syllable rates vary with temperature, researchers measured rates during the sustained portion of each call and related those measurements to temperature. The temperature relationships distinguished at least four song types among the sampled populations.

Males from northern Georgia and northern Alabama had the fastest rates, approximately 13.1 syllables per second at 25°C, corresponding to the newly described A. tallapoosa. Florida populations produced intermediate rates of about 10.0 syllables per second at the same temperature and retained the name A. bartrami. Males from western Alabama also sang at rates similar to those in Florida.

South Carolina populations had the slowest rates, approximately 5.8 syllables per second at 25°C, and were assigned to A. carolina. North Carolina males produced a distinctive paired pattern: the rate within syllable doublets was about 11.6 syllables per second at 25°C, while the rate between doublets was approximately 4.0 syllables per second. Call structure provided additional distinctions.

Florida A. bartrami males generally began a song with several shorter series of roughly five or six syllables before producing a sustained main section that averaged 107±22 syllables. The main series of A. carolina contained about 25 syllables, while A. peedee usually had about 17 and A. tallapoosa about 8. North Carolina males almost always arranged their syllables in doublets: of 517 syllables recorded in 30 main series from eight males, 482 belonged to doublets, eight were single syllables and nine occurred as triplets.

Spectrally, the calls were broadband, with most energy between 10 and 15 kilohertz, but cluster and principal-coordinate analyses detected significant population-level differences over both 30-second calls and individual syllables. Morphological measurements supported some of the divisions, although samples were uneven and several female characters did not differ significantly. Males assigned to A. tallapoosa were generally the smallest: their mean pronotal length was 5.08±0.14 millimetres, tegminal length 25.4±0.70 millimetres, hind-femur length 23.3±0.11 millimetres and hind-tibia length 25.1±0.20 millimetres.

A. carolina females had a mean pronotal length of 6.60±0.07 millimetres, compared with 5.99±0.23 millimetres for the Georgia and northern Alabama group. South Carolina A. carolina males had longer tegmina and greater hindwing exposure than A. parvipennis males, with respective means of 26.2±0.44 versus 23.8±0.47 millimetres for tegminal length and 2.74±0.17 versus 0.89±0.34 millimetres for hindwing exposure. The genetic analysis used mitochondrial cytochrome c oxidase subunit I, a roughly 3,700-base-pair ribosomal region containing ITS1, ITS2 and 5.8S, and the nuclear genes wingless and tubulin-alpha I.

Histone 3 showed no variation among the samples and was excluded from subsequent analyses. Bayesian multi-locus coalescent analyses indicated divergence among the sampled populations. The mitochondrial and ribosomal gene trees generally resembled the population tree, whereas the tubulin-alpha I tree differed substantially; the species-tree topology was unchanged when tubulin sequences were omitted.

Genetic relationships did not always coincide perfectly with broad song identity. Within South Carolina, the Georgetown County population grouped more closely with the North Carolina population in the geographic analysis, while the Aiken and Edgefield populations were genetically distinct from one another despite having the same general South Carolina song type. The authors say that additional sampling is needed to explain this variation.

More broadly, they propose that the flightless katydids’ limited movement, major river drainages and the fragmentation of longleaf pine habitat have reduced gene flow, allowing genetic divergence and song differentiation among populations. That explanation remains a hypothesis to be tested with broader geographic sampling rather than a directly measured effect of any particular river or habitat change. The three names also reflect the geographic structure recognized in the study.

A. carolina was named for South Carolina populations between the Savannah and Pee Dee rivers, A. peedee for populations north of the Pee Dee River in North Carolina, and A. tallapoosa for the area bounded by the Coosa and Tallapoosa rivers in Alabama. The researchers identified specific questions for future work: sampling both sides of the Pee Dee, Savannah, Coosa and Tallapoosa rivers could clarify the degree of isolation, while additional South Carolina populations could test why A. carolina displays marked genetic differentiation among sites. Their results nevertheless show that calling-song differences can reveal biodiversity overlooked by morphology alone, and that geographic, acoustic and genetic patterns together provide the basis for recognizing three new species in a previously uniform-looking katydid complex.

Subject of Research: Taxonomic and phylogeographic analysis of cryptic round-winged katydid species in the southeastern United States

Article Title: Geographic variation in the calling songs and genetics of Bartram’s round-winged katydid Amblycorypha bartrami ( Tettigoniidae , Phaneropterinae ) reveal new species

Article References: Forrest, T. G., Scobie, M., Brueckner, O., Bintz, B., & Spooner, J. D. (2023). Geographic variation in the calling songs and genetics of Bartram’s round-winged katydid Amblycorypha bartrami (Tettigoniidae, Phaneropterinae) reveal new species. Journal of Orthoptera Research, 32(2), 153-170. https://doi.org/10.3897/jor.32.96295

Image Credits: AI Generated

DOI: 10.3897/jor.32.96295

Keywords: cryptic species, calling songs, katydids, Amblycorypha, bioacoustics, phylogeography, coalescent analysis, flightless insects, southeastern United States, longleaf pine sandhills, Geographic, variation

The new species were found during macrofungal diversity surveys in mixed coniferous and broad-leaved forests containing Cupressaceae, Pinaceae, Theaceae and Fagaceae. Such forests are major habitat for Russula, whose species typically form ectomycorrhizal associations with trees. These partnerships involve fungal networks in plant roots and are described in the study as important for plant stress tolerance, root proliferation and nutrient acquisition. The discoveries therefore expand the documented diversity of ecologically important forest fungi, although the research did not directly test the functions of the three new species in living trees.

Russula is one of the most species-rich fungal genera worldwide, with an estimated minimum of 2,000 accepted species, and its members occur in ecosystems ranging from Arctic tundra to tropical forests. Many species resemble one another under the naked eye, and the genus is also known for substantial variation in features such as cap colour. The authors note that earlier morphology-based identifications can consequently obscure cryptic lineages and lead to misidentification. Molecular systematics has become essential for resolving these cases, particularly within Cyanoxanthinae, a group within the subgenus Heterophyllidiae.

The subsection was assigned to the three new taxa through a combination of anatomical and chemical characters: an inamyloid suprahilar region on the basidiospores, a metachromatic reaction of the pileipellis in cresyl blue, and unicellular pileocystidia. For each specimen, the researchers measured at least 40 basidiospores, 20 basidia and 20 cystidia. They also recorded the dimensions and ornamentation of spores, examined cystidia and pileipellis tissues, and used scanning electron microscopy to illustrate spore surfaces. Fresh collections were photographed and described in the field, while dried specimens were deposited in the Fungal Herbarium of Jiangxi Agricultural University.

Genetic evidence came from four loci: the internal transcribed spacer region, nuclear large subunit, mitochondrial small subunit and RNA polymerase beta subunit second largest subunit, or rpb2. The analysis contained 2,713 aligned characters from 57 taxa, including 372 parsimony-informative sites. Maximum likelihood and Bayesian inference analyses placed all three new species within Clade 1 of Cyanoxanthinae, and each formed an independent branch. Russula wuyishanensis and Russula purpureogrisea received complete support for their individual lineages, with ultrafast bootstrap values of 100% and Bayesian posterior probabilities of 1. Russula subpallescens grouped with Russula perviridis, but that broader relationship had unstable statistical support.

Russula purpureogrisea, whose name refers to its purplish-grey cap, is characterised by small to medium-sized fruiting bodies with a dark greyish-ruby to greyish-brown pileus. Its cap measures 20–50 mm across, and its crowded, unforked gills number 14–15 per centimetre near the margin. The spores measure 6.1–8.4 by 5.2–6.8 micrometres and bear mostly isolated, obtuse-conical warts, occasionally joined by fine lines or low ridges that sometimes form a local subreticulum. The authors distinguished it from related taxa through its pileus colour, spore ornamentation, cystidial dimensions and other structural details.

Russula subpallescens has a pinkish-white cap about 55 mm in diameter, crowded and equal-length gills with intervenose veins, and a hollow white stipe that develops pale yellow to maize-yellow patches with age or prolonged bruising. Its spores measure 6.7–8.4 by 5.8–6.8 micrometres, with dispersed amyloid warts, 0.3–0.8 micrometres high, frequently connected by short lines to produce a nearly subreticulate pattern. The species occupied a mixed coniferous and broad-leaved forest at an elevation of 1,526 metres in Xingshan County, Hubei. Its closest phylogenetic associate, R. perviridis, differs in having a greenish cap, reticulate spore ornamentation and frequently forked gills.

Russula wuyishanensis was named for Wuyishan, where the type specimen was collected in Fujian. It has a 45–75 mm reddish-grey to purplish-brown cap, sometimes with a greyish to dark greyish-green centre, and decurrent gills containing three types of shorter lamellulae. The context turns yellowish-brown when bruised. Its spores measure 6.2–9.7 by 5.2–7.7 micrometres and carry many obtuse warts that are sometimes connected by short lines or ridges and occasionally form a subreticulum. Three Fujian collections supported the description, with the holotype collected at 478 metres above sea level in 2023.

The findings also expose a limitation in a proposed visual division of Cyanoxanthinae. Earlier work divided the subsection into two major clades partly according to whether connections between spore warts were common or predominantly absent. Russula subpallescens fits that pattern because its warts are frequently joined by short lines. Russula purpureogrisea and R. wuyishanensis nevertheless clustered in the same molecular clade even though their spore ornamentation consists mainly of isolated warts with only partial or occasional connections. The authors therefore conclude that the boundary between the two clades is more complex than a simple count of spore-wart connections can indicate. Broader geographic sampling is needed to clarify the evolutionary history, ecology and full species diversity of the subsection in subtropical China.

Subject of Research: Taxonomic discovery of three Russula subsection Cyanoxanthinae species in subtropical China

Article Title: Three new species of Russula subsect. Cyanoxanthinae from subtropical China ( Russulales , Russulaceae )

Article References: Zhong, B.-J., Zeng, H., Wang, S.-N., & Yan, J.-Q. (2026). Three new species of Russula subsect. Cyanoxanthinae from subtropical China (Russulales, Russulaceae). MycoKeys, 134, 291-312. https://doi.org/10.3897/mycokeys.134.195117

Image Credits: AI Generated

DOI: 10.3897/mycokeys.134.195117

Keywords: Russula, Cyanoxanthinae, fungal taxonomy, mushroom diversity, ectomycorrhizal fungi, subtropical forests, phylogenetics, Hubei, Fujian, cryptic species, Three, species