First description on the female of Crambus duospineus Li, 2020 (Lepidoptera: Crambidae)

A majority of the known Colias species (Lepidoptera: Pieridae, Coliadinae) occur in the mountainous regions of Central-Asia, vast areas that are hard to access, rendering the knowledge of many species limited due to the lack of extensive sampling. Two gene regions, the mitochondrial COI ‘barcode’ region and the nuclear ribosomal protein RpS2 gene region were used for exploring the utility of these DNA markers for species identification. A comprehensive sampling of COI barcodes for Central Asian Colias butterflies showed that the barcodes facilitated identification of most of the included species. Phylogenetic reconstruction based on parsimony and Neighbour-Joining recovered most species as monophyletic entities. For the RpS2 gene region species-specific sequences were registered for some of the included Colias spp. Nevertheless, this gene region was not deemed useful as additional molecular ‘barcode’. A parsimony analysis of the combined COI and RpS2 data did not support the current subgeneric classification based on morphological characteristics.

Paraschistura alta , formerly reported from the eastern part of Sistan basin in Afghanistan, is reported for the first time from the Zahak River in Iranian part of this basin and its morphological characteristics, COI barcode and its phylogenetic relationship within other members of the genus Paraschistura from Iran are provided. All the measured meristic and morphometric characters of seven collected specimens of P. alta (SL= 65.8-112.1mm) from the Zahak River were in the range of its original description. The results of phylogenetic analysis based on the COI barcode region places the Iranian members of the genus Paraschistura into 11 groups, which show between 3.05% and 14.10% K2P sequence divergence. Paraschistura alta from Zahak River corresponds to a distinct clade, sister to P. bampurensis + P. hurmozensis .

The combination of morphological characteristics and DNA barcodes was used to a systematic study of Hippocampus spinosissimus,laying the foundation for rapid and accurate identification for the medical seahorse species. According to the reported literature and observation on seahorse samples,the typical characteristics of the H. spinosissimus include highly developed spiny,much short nose,single or double cheeks and strongly developed spines bordering pouch. Genomic DNAs of H. spinosissimus and other related seahorse species were extracted using the TIANamp Marine Animals DNA Kit. The COⅠ and ATP6 genes were amplified and sequenced in both directions. After the verification by Blast,the GC content,intraspecific and interspecific genetic distance,and the Neighbor joining( NJ) phylogenetic trees were analyzed by MEGA 7. The lengths of the COⅠ and ATP6 genes were 649 bp and 602-603 bp,respectively,with the average GC content of 39. 96% and 35. 37%. The maximum intraspecific genetic distances in H. spinosissimus based on COⅠ and ATP were both far less than the minimum interspecific genetic distance between H. spinosissimus and other seahorses,suggesting a significant barcoding gap. NJ analysis results of COⅠ and ATP6 exhibited that all H. spinosissimus species clustered together,indicating that the two DNA barcode could identify H. spinosissimus from other seahorses accurately and quickly. In addition,H. spinosissimus shared a close genetic relationship between H. kelloggi according to the NJ tree. Furthermore,there exits three stable subgroup structure of H. spinosissimus,indicating that COⅠ and ATP6 barcodes could be applied the indicator for the geographical ecology research of H. spinosissimus. The results obtained the typical morphological and molecular identification characteristics of H. spinosissimus,which played central roles for the development of species identification. This study provides an important basis data for expanding the medical seahorse resources and ensuring the safety of clinical medicine.

BackgroundThelazia callipaeda is a zoonotic parasitic nematode of the family Thelaziidae, with Phortica okadai as its intermediate host and only confirmed vector in China. China has the largest number of human cases of thelaziosis in the world. It is generally believed that infected domestic animals (dogs and cats) are the most important reservoir hosts of T. callipaeda, and thus pose a direct threat to humans. At present, there is little research or attention focused on the role of wildlife in the transmission cycle of thelaziosis in nature reserves.MethodsWe selected locations in four national nature reserves across China to monitor P. okadai and wildlife. We used a fly-trap method to monitor P. okadai density. Morphological analysis of the parasites collected from the conjunctival sac of the infected wildlife was undertaken as the first step in species identification, and polymerase chain reaction (PCR) was used for species confirmation.ResultsIn 2019, the density of P. okadai in Foping National Nature Reserve in China increased sharply, and infected P. okadai were newly found in the reserve. Giant panda, wild boar, leopard cat, and black bear were found to be newly infected with T. callipaeda (one individual of each species). A total of four worms were collected, one from each species of wildlife. The four worms were identified as T. callipaeda by their morphological characteristics; species identification was confirmed by PCR amplification.ConclusionsTo the best of our knowledge, this is the first report of T. callipaeda infection in P. okadai as well as in a variety of wildlife, including giant panda, in nature reserves in China. These results indicate that there is a transmission cycle of T. callipaeda among wildlife in these nature reserves. The increasing number of case reports of thelaziosis in wildlife suggest a likely risk of T. callipaeda infection for the inhabitants of villages situated around nature reserves.

Effective species conservation necessitates the ability to accurately differentiate among species, a challenge compounded by taxonomic uncertainties in freshwater mussels due to substantial intraspecific variation and pronounced phenotypic plasticity in shell morphology. The taxonomic status and species validity of Scabies longata and S. chinensis, two species endemic in China, have been under continuous debate since establishment. The lack of essential molecular data required for a comprehensive systematic study has resulted in the unresolved taxonomic status of these two species. This study presents molecular data, including COI barcoding, COI+28S rRNA, and mitogenomic data combined with morphological characteristics to assess the validity of S. longata and S. chinensis. Both morphological and COI barcoding data support the conclusion that S. longata and S. chinensis are junior synonyms of Nodularia douglasiae and N. nuxpersicae respectively. Our findings suggest the absence of Scabies species in China. Mitochondrial phylogenetic analyses were used to further elucidate intrageneric relationships within the genus Nodularia, revealing the following relationships: (N. breviconcha (Nodularia sp. 1 (N. douglasiae (N. nuxpersicae, N. nipponensis)))). We underscore the significance of employing an integrated taxonomic approach for species identification, especially given the considerable morphological disparities between larvae and adult freshwater mussels. Proper morphological identification of adult specimens is essential for extracting meaningful taxonomic characters. Furthermore, our findings suggest a notable resemblance between the freshwater bivalve fauna in southern China and those east of the Mekong River. ZooBank: urn:lsid:zoobank.org:pub:DA87D330-5E23-4F4B-8CC2-CBA3CD191BE8.

Mitochondrial COI barcoding of Pulicidae fleas and ultrastructural differentiation of the cat flea by scanning electron microscopy.

Little is known of the diversity of the monogenean parasites infesting deep-sea groupers, and there is even less information available about their geographic distributions within the ranges of their hosts. To improve our understanding of these host-parasite relationships we conducted parasitological evaluations of the deep-water Haifa grouper Hyporthodus haifensis from the southern Mediterranean off Tunisia and Libya. We collected more than one species of diplectanid monogeneans from this host, but among these only one dominant species was abundant. This proved to be morphologically very similar to Pseudorhabdosynochus sulamericanus Santos, Buchmann & Gibson, 2000, a species originally described from the congeneric host H. niveatus off Brazil and also recorded from H. niveatus and H. nigritus off Florida. Here, we conducted a morphological comparison between newly collected specimens and those previously deposited in museum collections by other authors. Further, we used COI barcoding to ascertain the specific identity of the three host species to better elucidate the circumstances that might explain the unexpectedly broad distribution of P. sulamericanus. We assigned our specimens from H. haifensis to P. sulamericanus primarily on the basis of morphological characteristics of the sclerotized vagina. We also noted morphological characteristics of eastern and western Atlantic specimens that are not clearly described or not given in previous descriptions and so prepared a redescription of the species. We confirmed, by COI barcoding, that no sister-species relationships were evident among the three hosts of P. sulamericanus. Our observation that P. sulamericanus infects unrelated host species with putatively allopatric distributions was unexpected given the very limited dispersive capabilities and the high degree of host specificity common to members of Pseudorhabdosynochus. This transatlantic distribution raises questions with regard to phylogeography and assumptions about the allopatry of Atlantic grouper species from the Americas and Afro-Eurasia. Here, we propose some hypothetical explanations for our findings.

BackgroundTaxonomy that utilizes morphological characteristics has been the gold standard method to identify mosquito species. However, morphological identification is challenging when the expertise is limited and external characters are damaged because of improper specimen handling. Therefore, we explored the applicability of mitochondrial cytochrome C oxidase subunit 1 (COI) gene-based DNA barcoding as an alternative tool to identify mosquito species. In the present study, we compared the morphological identification of mosquito specimens with their differentiation based on COI barcode, in order to establish a more reliable identification system for mosquito species found in Singapore.MethodsWe analysed 128 adult mosquito specimens, belonging to 45 species of 13 genera. Phylogenetic trees were constructed for Aedes, Anopheles, Culex and other genera of mosquitoes and the distinctive clustering of different species was compared with their taxonomic identity.ResultsThe COI-based DNA barcoding achieved a 100% success rate in identifying the mosquito species. We also report COI barcode sequences of 16 mosquito species which were not available previously in sequence databases.ConclusionsOur study utilised for the first time DNA barcoding to identify mosquito species in Singapore. COI-based DNA barcoding is a useful tool to complement taxonomy-based identification of mosquito species.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-014-0569-4) contains supplementary material, which is available to authorized users.

First Report of Leaf Spot Caused by Penicillium sclerotiorum on Aegiceras corniculatum in China.

Giant pandas have formed a series of strategies to adapt to bamboo food in the evolution process, such as eating bamboos at different altitudes in different seasons, and bamboos at different altitudes have specific morphological and environmental characteristics. In this study, Liziping National Nature Reserve in Sichuan province was taken as the study area, and the morphological characteristics and site environment differences of different staple food bamboos of giant panda populations in Xiaoxiangling Mountains were revealed through field investigation. The results showed that the basal diameter and height of A. spanostachya, Y. lineolata and F. ferax were significantly different. The F. ferax at low altitude had the largest basal diameter and height, while the A. spanostachya at higher altitude had the smallest basal diameter and height. Different bamboo species have different site environments, mainly in altitude and tree canopy density. Different bamboo species require different tree canopy which is most suitable for growth. The results of this study can provide reference for the habitat restoration and management of the giant pandas in this area, especially the refined management of the giant panda’s staple food bamboo.

Endophytic fungi were isolated from Macleaya cordata growing in Dabie Mountain by agar-block method, and then the endophytic fungi were grouped into different types based on their morphological characteristics, and thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) were employed to determine whether the metabolic substances contained sanguinarine or not, and then preliminarily identified by morphological method. The results showed that the leaves hosted the largest number of endophytes (96 isolates) followed by the stems (57 isolates) and finally the roots (28 isolates), respectively. Based on morphological characteristics the endophytic fungi were grouped into 26 types in our study. TLC and HPLC results showed that there was sanguinarine in the metabolic substances of BLH 51 strain. According to the morphological characteristic, the BLH 51 strain was identified as Fusarium proliferatum. All these indicated that the medicinal plant M. cordata harbors abundant endophytes, which could be a new source for the search of active secondary metabolites.

Accurate identification of species is fundamental for biological research and necessary for species conservation. DNA barcoding is particularly useful when identification using morphological characteristics is laborious and/or unreliable. However, barcodes for species are dependent on the availability of reference sequences from correctly identified specimens. The traditional use of morphology to delimit the species boundaries of Finnish bagworm moths (Lepidoptera: Psychidae: Naryciinae: Dahliciini) is controversial because there is overlap in their morphological characteristics. In addition, there are no suitable molecular markers. We verified the delimitation of seven out of eight previously described taxa, by using the currently standardized COI barcode and phylogenetic inference based on fragments of mitochondrial (COI) and nuclear genes (MDH). Moreover, we compared the results of molecular methods with the outcome of geometric morphometrics. Based on molecular identification, our findings indicate that there are five sexual species (Dahlica and Siederia spp.) and two parthenogenetic species (D. fennicella and D. triquetrella) in Finland. We suggest that molecular methods, together with geometric morphometrics of male genitalia are an effective way of delimiting species of bagworm moths.

Nature reserves are widely considered as one available strategy for protecting biodiversity, which is threatened by habitat fragmentation, and wildlife extinction. The Chinese government has established a goal of protecting 15% of its land area by 2015. We quantitated the characteristics and distribution of nature reserves in mainland China and evaluated the expansion process for national nature reserves. National nature reserves occupy 64.15% of the total area of nature reserves. Steppe and meadow ecosystem, ocean and seacoast ecosystem, and wild plant nature reserves represent lower percentages, particularly in national nature reserves, in which they comprised 0.76%, 0.54%, and 0.69%, respectively, of the area. Furthermore, medium and small nature reserves compose 92.32% of all nature reserves. The land area under any legal protection has reached 14.80%, although only 9.78% is strictly protected. However, if 9 super-large national nature reserves, located in Southwest and Northwest China were removed, the percentage of strictly protected area decreases to 2.66% of the land area of China. The areas contained in nature reserves in each province are not proportional to the areas of the provinces, particularly for national nature reserves, with higher protection rates in Southwest and Northwest China than in other regions. Of the 31 provinces, 22 provinces feature strict protection of less than 4% of their areas by national nature reserves; these provinces are mainly located in East, Central, South, and North China. Moreover, the unevenness indexes of the distribution of nature reserves and national nature reserves are 0.39 and 0.58, respectively. The construction of nature reserves has entered a steady development stage after a period of rapid expansion in mainland China. In recent years, the total area of national nature reserves has increased slowly, while the total area of nature reserves has not increased, although the number of nature reserves continues increase.

Fermentation processes using sanguinarine-producing fungi other than Macleaya cordata may be an alternative way to produce sanguinarine (SA), which is a quaternary benzo[c]phenanthridine alkaloid possessing antibacterial, anthelmintic, and anti-inflammatory properties. In this study, a SA-producing endophytic fungus strain BLH51 was isolated from the leaves of M. cordata grown in the Dabie Mountain, China. Strain BLH51 produced SA when grown in potato dextrose liquid medium. The amount of SA produced by this endophytic fungus was quantified to be 178 μg/L by HPLC, substantially lower than that produced by the host tissue. The fungal SA--which was analyzed by thin layer chromatography and high-performance liquid chromatography--was shown to be identical to authentic SA. Strain BLH51 was identified as Fusarium proliferatum based on the morphological characteristics and nuclear ribosomal DNA ITS sequence analysis. To the best of our knowledge, this is the first report concerning the isolation and identification of endophytic SA-producing fungi from the host plant, which further proved that endophytic fungi are valuable reservoirs of bioactive compounds.

China's national nature reserve network shows great imbalances in conserving the country's mega-diverse vegetation