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Southeast Asia and southern China (SEA-SC) harbor a highly diverse and endemic flora and fauna that is under increasing threat. An understanding of the biogeographical history and drivers of this diversity is lacking, especially in some of the most diverse and threatened groups. The Asian leaf-litter frog genus Leptolalax Dubois 1980 is a forest-dependent genus distributed throughout SEA-SC, making it an ideal study group to examine specific biogeographic hypotheses. In addition, the diversity of this genus remains poorly understood, and the phylogenetic relationships among species of Leptolalax and closely related Leptobrachella Smith 1928 remain unclear. Herein, we evaluate species-level diversity based on 48 of the 53 described species from throughout the distribution of Leptolalax. Molecular analyses reveal many undescribed species, mostly in southern China and Indochina. Our well-resolved phylogeny based on multiple nuclear DNA markers shows that Leptolalax is not monophyletic with respect to Leptobrachella and, thus, we assign the former to being a junior synonym of the latter. Similarly, analyses reject monophyly of the two subgenera of Leptolalax. The diversification pattern of the group is complex, involving a high degree of sympatry and prevalence of microendemic species. Northern Sundaland (Borneo) and eastern Indochina (Vietnam) appear to have played pivotal roles as geographical centers of diversification, and paleoclimatic changes and tectonic movements seem to have driven the major divergence of clades. Analyses fail to reject an "upstream" colonization hypothesis, and, thus, the genus appears to have originated in Sundaland and then colonized mainland Asia. Our results reveal that both vicariance and dispersal are responsible for current distribution patterns in the genus.
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Anuros/classificação , Biodiversidade , Filogenia , Animais , Ásia , Sequência de Bases , Teorema de Bayes , Filogeografia , Especificidade da Espécie , Fatores de TempoRESUMO
The horned toad assemblage, genus Megophrys sensu lato, currently includes three groups previously recognized as the genera Atympanophrys, Xenophrys and Megophrys sensu stricto. The taxonomic status and species composition of the three groups remain controversial due to conflicting phenotypic analyses and insufficient phylogenetic reconstruction; likewise, the position of the monotypic Borneophrys remains uncertain with respect to the horned toads. Further, the diversity of the horned toads remains poorly understood, especially for widespread species. Herein, we evaluate species-level diversity based on 45 of the 57 described species from throughout southern China, Southeast Asia and the Himalayas using Bayesian inference trees and the Generalized Mixed Yule Coalescent (GMYC) approach. We estimate the phylogeny using both mitochondrial and nuclear DNA data. Analyses reveal statistically significant mito-nuclear discordance. All analyses resolve paraphyly for horned toads involving multiple strongly supported clades. These clades correspond with geography. We resurrect the genera Atympanophrys and Xenophrys from the synonymy of Megophrys to eliminate paraphyly of Megophrys s.l. and to account for the morphological, molecular and biogeographic differences among these groups, but we also provide an alternative option. Our study suggests that Borneophrys is junior synonym of Megophrys sensu stricto. We provide an estimation of timeframe for the horned toads. The mitochondrial and nuclear trees indicate the presence of many putative undescribed species. Widespread species, such as Xenophrys major and X. minor, likely have dramatically underestimated diversity. The integration of morphological and molecular evidence can validate this discovery. Montane forest dynamics appear to play a significant role in driving diversification of horned toads.
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Anuros/classificação , Animais , Anuros/genética , Teorema de Bayes , Bufonidae/classificação , Bufonidae/genética , China , DNA/química , DNA/isolamento & purificação , DNA/metabolismo , DNA Mitocondrial/classificação , DNA Mitocondrial/genética , Filogenia , Filogeografia , RNA Ribossômico 16S/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
BACKGROUND: X-linked agammaglobulinemia (XLA) is a primary immune deficiency characterized by recurrent bacterial infections and profoundly depressed serum immunoglobulin levels and circulating mature B cells. It is caused by mutations of the Bruton tyrosine kinase (BTK) gene and is the most common form of inherited antibody deficiency. To our knowledge, this is the first report of XLA from Vietnam. METHODS: We investigated the BTK gene mutations and clinical features of four unrelated Vietnamese children. RESULTS: The mean ages at onset and at diagnosis were 2.5 and 8 years, respectively. All patients had a medical history of otitis media, pneumonia, and septicemia at the time of diagnosis. Other infections reported included sinusitis, bronchiectasis, arthritis, skin infections, meningitis, and recurrent diarrhea. We identified one previously reported mutation (c.441G >A) and three novel mutations: two frameshifts (c.1770delG and c.1742 delG), and one nonsense (c.1249A >T). CONCLUSIONS: The delayed diagnosis may be attributable to insufficient awareness of this rare disease on the background of frequent infections even in the immunocompetent pediatric population in Vietnam. Our results further support the importance of molecular genetic testing in diagnosis of XLA.
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Agamaglobulinemia/diagnóstico , Agamaglobulinemia/genética , Códon sem Sentido , Mutação da Fase de Leitura , Doenças Genéticas Ligadas ao Cromossomo X/diagnóstico , Doenças Genéticas Ligadas ao Cromossomo X/genética , Proteínas Tirosina Quinases/genética , Tirosina Quinase da Agamaglobulinemia , Artrite/complicações , Linfócitos B/metabolismo , Bronquiectasia/complicações , Relação CD4-CD8 , Criança , Pré-Escolar , Diarreia/complicações , Humanos , Masculino , Meningite/complicações , Neutropenia/etiologia , Otite Média/complicações , Pneumonia/complicações , Sepse/complicações , Sinusite/complicações , Dermatopatias Infecciosas/complicações , VietnãRESUMO
Species of bent-toed gecko (Cyrtodactylus) in Vietnam have been described at a rate of nearly four species per year since 2007 mostly based on morphological data. A tool that guides species delimitation will accelerate the rate of documentation, and at a time when the recognition of species greatly benefits conservation. We use DNA barcoding using COI (550 bp) to re-examine the levels of genetic divergence and taxonomic status of 21 described species of Vietnamese bent-toed geckos. Tree-based analyses resolve all sampled species and identify potential undescribed taxa. Kimura 2-parameter genetic distances between the described species average 21.0±4.2% and range from 4.3% to 28.7%. Further, our analyses discover two potentially new species from Vietnam, two from Laos and one from China. Herein we describe the new species Cyrtodactylus puhuensis sp. nov. from Vietnam on the basis of both genetics and morphology. Genetically, it differs from the remaining species by an average K2P distance of 24.0±1.8%. Morphologically, the new species is diagnosed by its medium-size (snout-vent length 79.24 mm and tail length 82.59 mm, for the single known individual), in having a series of moderately enlarged transverse subcaudals and a series of moderately enlarged femoral scales that extend from precloacal scales, in possessing femoral scales without pores, with males having five precloacal pores, and in exhibiting 8 supralabials, 10 infralabials, 23 narrow subdigital lamellae on its fourth toe, and 36 transverse ventrals.
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Lagartos/classificação , Lagartos/genética , Distribuição Animal , Estruturas Animais/anatomia & histologia , Animais , China , Código de Barras de DNA Taxonômico , Ecossistema , Feminino , Deriva Genética , Laos , Lagartos/anatomia & histologia , Masculino , Dados de Sequência Molecular , Filogenia , VietnãRESUMO
The number of described species of bent-toed geckos of the Cyrtodactylus irregularis species complex in Vietnam has increased from one to eight in the last six years. We combined morphological and molecular analyses to explore phylogenetic relationships among all described species in the group. The phylogeny required the description of two new species, Cyrtodactylus phuocbinhensis sp. nov. and Cyrtodactylus taynguyenensis sp. nov. Further, the tree resolved two additional undescribed clades that may also be new species. The species C. bugiamapensis and C. ziegleri were found to require redefinition. Cyrtodactylus phuocbinhensis sp. nov. is characterized by a series of enlarged femoral scales separated from preanal scales while Cyrtodactylus taynguyenensis sp. nov. does not possess enlarged femoral scales. Both new species are distinguished from other congeners by a combination of the following characters: small subcaudal scales, not transversely enlarged; presence (C. phuocbinhensis sp. nov.) or absence (C. taynguyenensis sp. nov.) of enlarged femoral scales; number of preanal pores; and dorsal pattern. Genetic distances between described species and new species were 16.5% and 2.0% in COI and RPL35, respectively, for C. phuocbinhensis sp. nov., and these distances were 18.8% and 2.2% for C. taynguyenensis sp. nov., respectively.
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Lagartos/genética , Animais , Lagartos/classificação , Filogenia , VietnãRESUMO
Introduction: Children are most often harmed by acute poisoning, which may cause disability or even death. This demonstrates the critical necessity for epidemiologic studies specific to each nation and area since they aid in developing plans for the prevention of acute poisoning. There are no data or outdated data on acute poisoning in children in Vietnam. This research would partly fill this existing gap and compare the trend with other places across the globe. Methods: A retrospective study was conducted in the 10-year period from 2012 to 2021 in Haiphong Children's Hospital, Vietnam. Results: There were 771 children hospitalized due to acute poisoning. Children in the 1-5-year-old group accounted for the highest rate, at 506 (65.6%). The mean age was 4.5 ± 4.1 years old. The male-to-female ratio was 1.2/1. Nonpharmaceutical chemicals were the most common agent in 331 cases (42.9%), including cleaning products 63 (19.0%), rat poison 60 (18.1%), and petrol 42 (12.7%). Medications were the second most common agent in 290 cases (37.6%), mostly paracetamol 60 (20.7%) and sedatives 40 (13.8%). There were 633 (82.1%) children exposed to poisons unintentionally. Conclusion: Children between the ages of 1 and 5 are more likely to be exposed to harmful substances. The most common agents were nonpharmaceutical chemicals followed by pharmaceuticals. Most incidents were inadvertent. Finally, our research may provide insights that public health authorities might use to plan practical actions.
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OBJECTIVES: To improve 4 skills (communication, history-taking, past history-taking, and documentation) in medical students, we designed and pilot-tested a curriculum to teach a sample of Year 4 (Y4) students these skills and compared the clinical performance of these students with students not receiving the intervention. METHODS: The study focused on the new curriculum's effectiveness in enhancing students' performance of these skills. To minimize exposure across groups, participants were divided into intervention and control groups at random and placed in various classrooms. We evaluated each group's clinical competency 3 times: prior to the intervention, 9 weeks afterward, and 2 years later. RESULTS: There was no difference at baseline between the 2 groups. Immediately following the intervention, the mean score of the intervention group's skills was significantly higher than before and higher than the control group in each clinical skill. The performance difference between the 2 groups was maintained for 2 years following the intervention. CONCLUSIONS: Following a 9-week curriculum, evaluators rated students' performance higher than their counterparts who learned these skills through standard informal exposure in the clinical setting. The fact that this performance advantage was maintained for 2 years following the intervention is a testament to the durability of the intervention and the value of dedicated training in these critical areas at an early point in students' clinical careers.
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We describe a new species of Dendrelaphis from the coastal area in southern Vietnam based on morphological data and nucleotide sequences from one partial mitochondrial gene (Cytb). Dendrelaphis binhi sp. nov. is diagnosed by the following morphological characters: body scale rows 13 at neck and midbody, 9-11 before vent; vertebral scale row feebly enlarged; ventrals 154-158 in males and 161-170 in females; subcaudals 95-106 in males and 95-102 in females; one supralabial touching the eye; 18 or 19 maxillary teeth; hemipenis spinose, reaching 10th or 11th SC, with a slender papilla; cloacal plate divided; dorsum dark brown; and a white stripe along the lower flank. The new species differs from its congeners by an uncorrected p-distance in Cytb sequences of at least 8.1%.
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Colubridae , Lagartos , Masculino , Feminino , Animais , Vietnã , Estruturas Animais/anatomia & histologia , Distribuição Animal , FilogeniaRESUMO
A new species of the genus Vietnamophryne is described from Vietnam on the basis of two specimens collected from Tuyen Quang Province, Northeastern Vietnam. The new species is morphologically most similar to Vietnamophryne occidentalis from Thailand, however, it differs from the latter by having large black blotches in the lower jaw region, and a yellow-orange chest and belly. The genetic distance between the new species and other Vietnamophryne taxa is > 2.13% (16S mtDNA gene fragment). Vietnamophryne aurantifusca sp. nov. can be distinguished from all other species of Vietnamophryne by a combination of the following morphological characteristics: Size medium (SVL 17.618.2 mm in males); head wider than long; tympanum medium; finger I longer than half of finger II; dorsal skin relatively smooth with some round nodules, concentrated in the middle of the back, arranged along the length of the back, with a prominent ridge along the spine; Dorsum orangish-brown entirely and paler on margin of back with a small brownish ridge along the spine; sides brownish with creamy patches and orange spots; ventral surface orange, with grey marbling, most intense on the throat, ventral side of arms and thighs, and ventral surfaces of limbs dark grey with some orange spots.
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Anuros , Tórax , Masculino , Animais , Vietnã , Filogenia , ExtremidadesRESUMO
We describe a new species of kukri snake from Da Lat City and its vicinity in Lam Dong Province, southern Vietnam on the basis of morphological characters of four specimens, including two males and two females. Oligodon tuani sp. nov. differs from other congeners by a combination of the following characters: large size in adults (TL ≤ 888 mm); 19 dorsal scale rows at midbody; ventrals 173-179 in males and 187-193 in females; subcaudals 58 or 59 in males and 44 or 45 in females; presubocular present; 8 supralabials, fourth and fifth entering orbit; cloacal plate undivided; 10 maxillary teeth, posterior three enlarged; vertebral stripe present; temporal streak present but faint and interrupted; long and deeply forked hemipenes, extending to 25th subcaudal, without spines and papillae; and 13-15+3-4 dorsal blotches.
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Colubridae , Lagartos , Feminino , Masculino , Animais , Vietnã , Estruturas Animais/anatomia & histologia , Distribuição Animal , FilogeniaRESUMO
Introduction: In individuals with urinary tract infections, Escherichia coli (E. coli) is an ubiquitous causative agent and antibiotic resistance is on the rise throughout the world. Therefore, early diagnosis and appropriate choice of antimicrobials are essential. The purpose of our study is to describe some of the clinical and epidemiological characteristics and the laboratory test results of children treated in our hospital for urinary tract infections caused by E. coli. Methods: The study included 128 patients from 2 months to 15 years of age with urinary tract infections caused by E. coli and treated at the Haiphong Children's Hospital during the periods of 2011-2013 and 2018-2020. Results: During the two study periods, 57 and 71 cases, respectively, were included. The most common clinical symptom was fever in 40 and 46 cases, respectively. The proportion of E. coli's resistance to ampicillin increased from 85.3% in 2011-2013 to 97.1% in 2018-2020. In 2011-2013, 70.5% of E. coli isolates were resistant to cotrimoxazole, which increased to 81.4% during 2018-2020. During both periods, E. coli was highly sensitive to amikacin, at 87% and 95.5%, respectively. In 2018-2020, carbapenems (meropenem and imipenem) and piperacillin were also effective against E. coli. Conclusion: Our study revealed that high fever was the most prevalent clinical characteristic in urinary tract infections caused by E. coli in children and E. coli was mostly resistant to ampicillin, nalidixic acid, and cotrimoxazole but was highly sensitive to ciprofloxacin, amikacin, piperacillin, meropenem, and imipenem.
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The congenital immune system includes neutrophils, which perform a variety of functions. Congenital and acquired neutropenia are rare illnesses with an underestimated prevalence in children. The aim of this study is to examine the epidemiology and etiology of febrile neutropenia in children at Haiphong Children's Hospital, Haiphong, Vietnam. Methods: A cross-sectional study was carried out on 421 febrile neutropenia children. Clinical and laboratory characteristics were examined. Results: The median age (IQR) was 25.0 (12.5-59.5) months. The male-to-female ratio was 1.35/1. There were twice as many children living in the suburbs (66.98%) as in urban areas (33.02%). The mean (SD) temperature at admission was 38.50 ± 0.59 °C. Diagnosed causes associated with neutropenia included acute respiratory infections 250 (59.45%), gastrointestinal infections 68 (16.1%), erythema 37 (8.79%), acute leukemia 15 (3.56%), urinary tract infection 5 (1.19%), and encephalitis/meningitis 4 (0.95%). Viral etiology accounted for 61.52% (259): influenza type A-50.19% (130), influenza type B-31.27% (81), dengue virus-14.67% (38), measles virus 1-93% (5), rotavirus-1.54% (4), and EBV-0.4% (1). Twenty-five patients (5.94%) were found to have bacteria in their cultures, with Streptococcus pneumonia being the most common (eight patients; 32%). Conclusions: Febrile neutropenia was common in children under 2 years old. Primary clinical manifestations were acute upper respiratory tract infections, and viruses most commonly caused febrile neutropenia. Further studies with larger sample sizes are needed to determine the cause of febrile neutropenia.
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INTRODUCTION: Bronchiolitis is the most prevalent cause of hospitalization in infants under the age of 12 months. The disease is caused by respiratory syncytial virus (RSV) infection, which can cause breathing difficulties and respiratory failure. Therefore, it is necessary to discover the risk factors of severe bronchiolitis to diagnose and treat promptly. This study is aimed at describing the epidemiological characteristics and clinical features of acute bronchiolitis caused by RSV and assessing the related factors to severe acute bronchiolitis in studied patients. METHODS: A descriptive cross-sectional study was carried out in Haiphong Children's Hospital, Haiphong, Vietnam, for one year, from October 1, 2016, to September 30, 2017. All bronchiolitis admissions < 2 years were included. RESULTS: 377 children were evaluated, including 261 boys and 116 girls; children under 6 months accounted for the highest proportion (57%), and 47 (12.5%) of all patients had severe disease. Wheezing was the main reason to be taken to the hospital 261 (69.2%). Clinical symptoms of acute bronchiolitis such as cough, tachypnea, and runny nose were found in all patients. Bronchiolitis cases increased in the winter-spring season, and the highest registered number of patients was 42 in March. Image of bronchiolitis on chest X-ray was found in all patients, and air trapping lung was found in 124 (32.9%) patients. The risk factors included age (≤6 months), low birth weight, preterm birth, nonbreastfeeding for the first six months, early weaning, and exposition to cigarette smoke increased the severe disease (p < 0.05). CONCLUSION: The number of hospitalized infants with bronchiolitis caused by RSV has an upward trend during the winter-spring season (from October to March). This study confirms that age, preterm birth, breastfeeding under 6 months, history of exposure to cigarette smoking, low birth weight, having sibling(s) under five years old going to kindergarten, history of undergoing cesarean section, history of mechanical ventilation, poor living condition, and maternal education are 10 risk factors of severe bronchiolitis caused by RSV.
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A new vine snake, genus Ahaetulla, from Soc Trang Province, southern Vietnam is described based on morphological data and nucleotide sequences from COI and Cytb. Ahaetulla rufusoculara sp. nov. is diagnosed by the following morphological characters: bright red eyes; snout without dermal appendage; internasal separated from supralabial by loreal; body scale rows 15-15-13; ventrals 186190 in males and 182185 in females; subcaudals 143153 in males and 128 or 129 in females; 8 (rarely 9) supralabials, fourth and fifth or fourth to sixth entering orbit; 1214 maxillary teeth; hemipenis short, reaching 6th or 7th SC; cloacal plate divided; dorsum bright green; and yellow or white stripe along the lower flank. The new species differs from its congeners by an uncorrected p-distance in COI and Cytb sequences of at least 7.7% and 7.5%, respectively.
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Colubridae/anatomia & histologia , Colubridae/classificação , Estruturas Animais , Animais , Feminino , Masculino , Filogenia , VietnãRESUMO
A new species of the genus Gekko Laurenti, 1768 is described from Phu Yen Province, southern Vietnam based on morphological and molecular evidence. Gekko phuyenensis sp. nov. differs from other congeners by its moderate size, with a snout-vent length of 90.292.8 mm; nares in contact with rostral; internasal 1; interorbitals 35 or 36; ventrals 3032; dorsal tubercle rows 9 or 10; precloacal pores 12 or 13 in males and precloacal pits 13 in female; subdigital lamellae under first finger 1821, fourth finger 19 or 20, first toe 1820, and fourth toe 2123; subcaudals median enlarged; no webbing between toes; and the absence of tubercles on hind limbs. Molecular phylogenetic analyses of mitochondrial gene COI (681 bp) using the geckos from Phu Yen and its congeners in southern Vietnam resolve the new species as the sister lineage to G. cf. grossmanni from Khanh Hoa Province. Genetic distances between G. phuyenensis sp. nov. and its congeners in southern Vietnam range from 16.924.2%. The discovery of this new species raises the number of known species of Gekko in Vietnam to 14.
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Lagartos/anatomia & histologia , Lagartos/classificação , Estruturas Animais , Animais , Feminino , Genes Mitocondriais , Masculino , Filogenia , VietnãRESUMO
We describe a new species of the genus Subdoluseps Freitas, Datta-Roy, Karanth, Grismer Siler from a coastal area in southern Vietnam. Subdoluseps vietnamensis sp. nov. is characterized by the following morphological characters: medium size in adults (snout-vent length up to 48.7 mm); tail length/snout-vent length ratio 1.04; toes not reaching finger when limbs adpressed; 27-30 midbody scale rows, smooth; 55-57 paravertebral scales; 55-62 ventral scale rows; 64-74 subcaudal scales; frontoparietal scale single; four supraoculars; prefrontals not in contact with one another; two loreal scales; seven supralabials; ear-opening with two lobules on the anterior border; smooth lamellae beneath finger III 9 or 10 and toe IV 12-15; six enlarged precloacal scales; and four distinct black stripes on dorsum. The new species differs genetically from its closest congeners, S. bowringii (Günther) and S. frontoparietale (Taylor), by uncorrected p-distances of 10.0% and 9.5%, respectively in ND1 sequences, and clusters into the same matriline with these two congeners on the phylogenetic trees.
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Lagartos , Distribuição Animal , Estruturas Animais , Animais , Ecossistema , Lagartos/classificação , Filogenia , VietnãRESUMO
King Cobra (Ophiophagus hannah) bite is well-known for its potentially fatal neurotoxicity. However, fatalities still occur, despite specific antivenom and respiratory support. Cardiovascular disturbances, which have attracted little attention in published reports of O. hannah envenoming, could contribute to fatality. We present two cases of confirmed O. hannah envenoming in Southern Vietnam in which there were cardiac abnormalities including arrhythmias and electrocardiographic changes, as well as elevated markers of myocardial damage. Cardiac pacing was required. One patient developed critical multi-organ dysfunctions partly explained by extensive necrotizing fasciitis/myositis originating from an Aeromonas sobria wound infection. This resulted in rhabdomyolysis, disseminated intravascular coagulation and acute kidney injury. Specific antivenom reversed neurotoxic effects of envenoming. Additional therapeutic interventions included antibiotics, surgical debridement, continuous renal replacement therapy and therapeutic plasma exchange. Both patients eventually made full recoveries. Apart from the critical problem of rapidly evolving and severe neurotoxicity, our case reports also emphasises the risk of cardiotoxic envenoming, and the complications of an overwhelming secondary bacterial wound infection. We suggest a practical approach to diagnosis and management.
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Fasciite Necrosante , Ophiophagus hannah , Aeromonas , Animais , Venenos Elapídicos , Humanos , VietnãRESUMO
A new skink of the genus Scincella Mittleman, 1950, from Binh Phuoc Province, southern Vietnam is described based on morphological data, including hemipenial characters, and nucleotide sequences from COI. The following morphological characters diagnose Scincella baraensis sp. nov.: relatively small size in adults (snout-vent length up to 49.2 mm); when limbs adpressed, toes do not or just reach fingers; 30 smooth midbody scale rows and dorsal scales not enlarged; 66-70 paravertebral scales; 64-66 ventral scale rows; 7 supralabials, the fifth and sixth below the center of the eye; enlarged temporal scales 1+2; nuchals present; ear with small auricular lobules; 18-20 smooth lamellae beneath toe IV; hemipenis smooth, short, and unforked; dorsum with faint black dots. The new species differs from its congeners by an uncorrected p-distance in COI sequences of at least 16.2%.
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Lagartos , Distribuição Animal , Estruturas Animais , Animais , Tamanho Corporal , Filogenia , VietnãRESUMO
A new species of Sinomicrurus Slowinski, Boundy, and Lawson, 2001 is described herein based on a series of specimens. The new species, Sinomicrurus peinani sp. nov., occurs in southern China and northern Vietnam. Sinomicrurus peinani sp. nov. is distinguished from its congeners by the following combination of characters: (1) 30-35 black cross-bands on body and tail; (2) 13 dorsal scale rows throughout, all smooth; (3) white belly with black cross-bands or irregular spots; (4) broad white transverse bar on top of head with inverted V-shaped anterior margin, white bar wider than anterior black bar; and (5) frontal V-like, 1.3 times as long as wide. In addition, new occurrences of S. houi in Guangxi, China, and Vietnam are discussed.