Antimicrobial peptide defenses of the Tarahumara frog,Rana tarahumarae

Populations of the Tarahumara frog Rana tarahumarae have decreased markedly in recent years in the northern part of their range. Infection by the chytrid fungus Batrachochytrium dendrobatidis has been implicated in these declines. To determine whether antimicrobial peptides in the skin provide protection against this pathogen, norepinephrine-stimulated skin secretions were tested for their ability to inhibit growth of B. dendrobatidis in vitro. After concentration, crude mixtures of skin peptides inhibited the growth of the chytrid in a concentration-dependent manner. Proteomic analysis led to the identification and characterization of three peptides belonging to the brevinin-1 family of antimicrobial peptides and three belonging to the ranatuerin-2 family. The two most abundant peptides, ranatuerin-2TRa (GIMDSIKGAAKEIAGHLLDNLKCKITGC) and brevinin-1TRa (FLPVIAGIAANVLPKLFCKLTKRC), were active against B. dendrobatidis (MIC of 50 microM for ranatuerin-2TRa and 12.5 microM for brevinin-1TRa against zoospores). These data clearly show that antimicrobial peptides in the skin secretions of the Tarahumara frog are active against B. dendrobatidis and should provide some protection against infection. Therefore, the observed susceptibility of these frogs to this pathogen in the wild may be due to the effects of additional environmental factors that impair this innate defense mechanism, leading to the observed population declines.     

Icosiella turgeocauda n. sp. (Nematoda: Onchocercidae) and Seuratascaris numidica (Nematoda: Ascarididae), parasites of the frog,Rana cancrivora (Anura: Ranidae), from Luzon,Republic of the Philippines

Icosiella turgeocauda n. sp. from the intestinal mesenteries of Rana cancrivora collected at Luzon, Republic of the Philippines, is described and illustrated. Icosiella turgeocauda n. sp. represents the ninth species to be assigned to the genus and is easily differentiated from all the previously described species by the position of the vulva and the presence of bilateral umbos on the caudal end of the male. Seuratascaris numidica also was found. The Philippines represents a new location record for S. numidica.     

New species of Oswaldocruzia (Nematoda: Molineoidae), new species of Rhabdias (Nematoda: Rhabdiasidae), and other helminths in Rana cf. forreri (Anura: Ranidae) from Costa Rica

Oswaldocruzia costaricensis n. sp. (Strongylida: Molineidae) from the intestines and Rhabdias savagei n. sp. (Rhabditida: Rhabdiasidae) from the lungs of Rana cf. forreri (Anura: Ranidae) are described and illustrated. Oswaldocruzia costaricensis represents the 77th species assigned to the genus and differs from the other Neotropical species in the genus by possessing a Type II bursa and long cervical alae. Rhabdias savagei represents the 47th species assigned to the genus and differs from other Neotropical species in the genus by possession of 4 lips and a postequatorial vulva. Rana cf. forreri was also found to harbor the trematodes, Haematoloechus parcivitellarius and Megalodiscus temperatus, the nematodes, Aplectana incerta, Aplectana itzocanensis, Cosmocerca podicipinus, Foleyellides striatus, Subulascaris falcaustriformis, and a larva of the nematode Brevimulticaecum sp. Cosmocerca panamaensis is considered to be a synonym of Cosmocerca podicipinus.     

Histology of the esophagus of the adult frog Rana perezi (Anura: Ranidae).

Study of the esophageal microscopic morphology of adult Rana perezi by light and electron microscopy discloses some large folds throughout the esophagus that are in themselves ringed. Glandular ostia open in the furrows of the luminal surface. The esophageal wall is made up of a connective adventitia rich in melanocytes, a muscular tunica, a connective and glandular subepithelial layer, and a pseudostratified ciliated epithelium. This epithelium basically consists of ciliated, goblet, basal, microvillous-apex, and migratory cells. Two types of goblet cells are distinguished with regard to the granular ultrastructure. The microvillous-apex cell has not been found in other amphibians. It shows a very differentiated morphology with a high number of mitochondria. The basal cells give the epithelium a pseudostratified morphology, and they have a proliferative function. Glands are branched and drain through an excretory duct that has a monolayered mucosecreting epithelium. The glandular units are formed by two principal types of cells: mucosecretory and serous.     

Growth of the pectoral girdle of the Leopard frog, Rana pipiens (Anura: Ranidae)

This article describes the growth of the anuran pectoral girdle of Rana pipiens and compares skeletal development of the shoulder to that of long bones. The pectoral girdle chondrifies as two halves, each adjacent to a developing humerus. In each, the scapula and coracoid form as single foci of condensed chondrocytes that fuse, creating a cartilaginous glenoid bridge articulating with the humerus. Based on histological sections, both the dermal clavicle and cleithrum begin to ossify at approximately the same time as the periosteum forms around the endochondral bones. The dermal and endochondral bones of the girdle form immobile joints with neighboring girdle elements; however, the cellular organization and growth pattern of the scapula and coracoid closely resemble those of a long bone. Similar to a long bone epiphysis, distal margins of both endochondral elements have zones of hyaline, stratified, and hypertrophic cartilages. As a result, fused elements of the girdle can grow without altering the glenoid articulation with the humerus. Comparisons of anuran long bone and pectoral girdle growth suggest that different bones can have similar histology and development regardless of adult morphology.     

Polystoma macrocnemis n. sp. (Monogenea: Polystomatidae) from the Iranian longlegged wood frog Rana macrocnemis (Ranidae) in Turkey

Polystoma macrocnemis n. sp. (Polystomatidae) is described from the urinary bladder of Rana macrocnemis (Ranidae) in Turkey. Its general morphology is similar to that of other members of the genus, but it is distinguished from closely related species by numerical parameters of the hamuli and also by its host species and geographical range.     

Systematics of a widespread Southeast Asian frog,Rana chalconota (Amphibia: Anura: Ranidae)

The abundant Sundaland forest frog, Rana chalconota, has long been considered a single widespread species, although some authors have recommended its division into regional subspecies. The discovery of co‐occurring pairs of morphologically distinct populations in three widely separated parts of the range led to a morphological and molecular analysis of populations from all parts of the known range. The results suggest that R. chalconota consists of at least seven species from Thailand through Borneo and Java. Existing names are applied to three of these species, R. chalconota (Schlegel), R. raniceps (Peters) and R. labialis Boulenger. We describe four others as new species and suggest the existence of one or two additional, unnamed species. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 155 , 123–147.     

Helminth infracommunities of Rana vaillanti brocchi (Anura: Ranidae) in Los Tuxtlas, Veracruz, Mexico

A total of 76 adult individuals of Rana vaillanti were collected in Laguna Escondida, Los Tuxtlas, Veracruz, Mexico, and their helminth infracommunity structure was determined. Among the 21 helminth taxa collected (10 digeneans, 8 nematodes, and 3 acanthocephalans), the digenean Langeronia macrocirra reached the highest prevalence (64.4%), mean abundance (6.6), and mean intensity (10.4), as well as the highest total number of individuals (499). Only 2 frogs were uninfected, the remainder harbored between 1 and 7 helminth species and 1-102 individuals; mean species richness and abundance were 3.49 +/- 0.22 and 16.1 +/- 16.3, respectively. Langeronia macrocirra dominated in 50.6% of the infracommunities, with relatively low Berger-Parker index values (0.56); for this reason, the evenness was high (0.70 +/- 0.31), and consequently, diversity values are the highest recorded to date in species of Rana. However, patterns of helminth infracommunity richness and diversity were similar to those previously observed in amphibians. This structure is attributed to the feeding habits (between 66.7 and 81% of helminth species parasitizing R. vaillanti enter using the food web dynamics) and low vagility (the remainder species infect by host penetration).     

Phylogeography of the fanged dicroglossine frog, Limnonectes fujianensis (Anura, Ranidae), in Taiwan

A phylogenetic analysis of Taiwanese fanged dicroglossine frog, Limnonectes fujianensis (Anura, Ranidae), was conducted to examine its genetic diversification using sequence data from a portion of the mitochondrial DNA (mtDNA) cytochrome b sequences. We collected genetic data from 200 individuals at 23 localities in Taiwan and three localities in China. A neighbor-joining tree of 39 haplotypes revealed two clades in Taiwan and a clade in China, each showing restricted geographical distribution. The pattern of geographical divergence suggests a single invasion into Taiwan. Divergence times between clades were inferred using molecular clock tests. The population relationship of L. fujianensis between Taiwan and mainland China, and the phylogenetic relationships with its congeners, e.g., L. bannaensis, L. fragilis and L. kuhlii, were obtained and discussed.     

Extra-intestinal localization of Goussia sp. (Apicomplexa) oocysts in Rana dalmatina (Anura: Ranidae), and the fate of infection after metamorphosis

Although coccidia of the genus Goussia are common parasites of fish, only 2 species have been described in amphibians: G. hyperolisi from common reed frogs Hyperolius viridiflavus from Kenya and G. neglecta from unspecified European water frogs of the genus Rana from Germany. The genus Goussia is characterized by an oocyst, with a fine oocyst wall, containing 4 dizoic sporocysts that are composed of 2 valves joined by a longitudinal suture and lacking a Stieda body (typical for the genus Eimeria). To date, infections in amphibians were generally considered to be specific to the intestine of aquatic larval stages (tadpoles) of anurans. Herein, we report on: (1) the presence of oocysts of Goussia sp. in an extra-intestinal location (liver) of tadpoles of the agile frog R. dalmatina and (2) the presence of oocysts in the liver of both juvenile and subadult R. dalmatina. These observations represent novel traits for Goussia infections in amphibians; they may explain the vertical transmission of Goussia in tadpoles.     

Strongyluris panamaensis n. sp. (Nematoda: Heterakidae) and other helminths from the lizard,Anolis biporcatus (Sauria: Polychrotidae), from Panama

Strongyluris panamaensis n. sp. from the intestine of Anolis biporcatus collected in Panama is described and illustrated. Strongyluris panamaensis represents the 30th species assigned to this genus and the fourth from the neotropical region. Strongyluris panamaensis can be easily distinguished from all other neotropical species because it has the shortest spicule. In addition to S. panamaensis, 3 species of Trematoda, Mesocoelium monas, Parallellopharynx arctus, and Urotrema scabridum, and 3 species of Nematoda, Crytosomum penneri, Oswaldocruzia bainae, and Piratuba digiticauda, were identified.     

Phylogeography of the rice frog, Fejervarya multistriata (Anura: Ranidae), from China based on mtDNA D-loop sequences

The rice frog, Fejervarya multistriata, is an amphibian widely distributed in China. In this study, we sampled the species across its distributional area in China and sequenced the mtDNA D-loop to investigate the genetic diversity and geographical pattern of the frog population. The results revealed 38 haplotypes in the population, with K2P values varying from 0.19% to 4.22%. Both a phylogenetic analysis and a nested clade analysis (NCA) detected two geographically isolated lineages respectively distributed around the Yangtze drainage (Yangtze lineage) and the south of China (southern lineage). NCA inferred a contiguous range expansion within the Yangtze lineage and allopatric fragmentation within the southern lineage, which might be partly due to the limited samples from this lineage. Accordingly, Fu's Fs test also indicated a population expansion after glacial movement. Therefore, we assumed that the species history responding to glacial events shaped the present population pattern of F. multistriata on the Chinese mainland.     

Cosmocerca vrcibradici n. sp. (Ascaridida: Cosmocercidae), Oswaldocruzia vitti n. sp. (Strongylida: Molineoidae), and other helminths from Prionodactylus eigenmanni and Prionodactylus oshaughnessyi (Sauria: Gymnophthalmidae) from Brazil and Ecuador

Cosmocerca vrcibradici n. sp. and Oswaldocruzia vitti n. sp., intestinal parasites of Prionodactylus eigenmanni (type host) and P. oshaughnessyi, are described and illustrated. Of the 19 valid species of Cosmocerca, C. vrcibradici n. sp. represents the 12th Neotropical species and the first species to be reported from lizard hosts. Of the 76 valid species of Oswaldocruzia, O. vitti n. sp. represents the 14th Neotropical species and the eighth species to be reported from lizard hosts. In addition, 1 species of Digenea, Mesocoelium monas, and 1 species of Acanthocephala, Acanthocephalus saurius, were found.     

New genus of Cosmocercidae (nematoda) and other helminths in Hylarana volkerjane (Anura: Ranidae) from papua new Guinea

Paraplesiohedruris rinse n. gen., n. sp. (Ascaridida; Cosmocercoidae; Cosmocercidae) from the large intestine of Hylarana volkerjane (Anura; Ranidae) is described and illustrated. The new genus is assigned to the Cosmocercinae of the Cosmocercidae based on the presence of an esophagus composed of a short pharynx, cylindrical corpus, isthmus, and valved bulb; on the presence in males of paired spicules and numerous caudal papillae; plus the presence in the female of an equatorial vulva, 2 uteri, a short tail, and thin-shelled eggs. The Cosmocercinae now contains 10 genera. Hylarana volkerjane was also found to harbor 6 additional species of Nematoda, adults of Aplectana macintoshii, Icosiella papuensis, Meteterakis crombiei, Paracapillaria spratti, Physalopteroides milnensis, and larvae of Abbreviata sp., as well as 1 species of Acanthocephala, Pseudoacanthocephalus bufonis.     

Paracosmocercella rosettae n. gen. et n. sp. (Nematoda: Cosmocercoidea: Cosmocercidae) collected from the Japanese tree frog, Hyla japonica (Anura: Hylidae), in Japan

A new genus, Paracosmocercella (Nematoda: Cosmocercoidea: Cosmocercidae), is proposed with monotypic Paracosmocercella rosettae n. sp. from the Japanese tree frog, Hyla japonica Günther, 1859, in Oita, Kyushu Island, Japan. Paracosmocercella resembles Cosmocercella and Cosmocercoides by having 2 rows of rosette papillae without plectanes ventrally in preanal region of male, but it is readily distinguished from Cosmocercella by lacking clear vesicle supporting rosette papillae and from Cosmocercoides by lacking rosette papillae in perianal region and having large-sized eggs, which hatch in uterus.     

Polystoma luohetong n. sp. (Monogenea: Polystomatidae) from Rana chaochiaoensis Liu (Amphibia: Ranidae) in China

Systematic Parasitology - Polystoma chaochiaoensis from the urinary bladder of the chaochiao frog Rana chaochiaoensis Liu was briefly described in a symposium abstract and presented at the Third...     

Population estimation in the Torrent frog, Amolops lamtensis (Anura: Ranidae)

The results of four attempts at yearly intervals to estimate populations of Amolops lamtensis (Blgr.) along 360 m of forest stream are discussed and the most consistent estimates are obtained using Jolly's (1965) method of analysing such data. Observations of movement within the population indicate that most frogs are recaptured close to the original point of capture although a few show considerable downstream movement, possibly as a result of being swept away by the current. The recapture expectations are not, however, random and there is a strong indication that larger frogs are more liable to recapture. There is also evidence that frogs are available for recapture for different lengths of time, and it is likely that this leads to an under-estimate of the total population.     

Haematoloechus danbrooksi n. sp. (Digenea: Plagiorchioidea) from Rana vaillanti from Los Tuxtlas,Veracruz, Mexico

Haematoloechus danbrooksi n. sp. from the lungs of Rana vaillanti in Veracruz state, Mexico, was found. The new species is most similar morphologically to H. medioplexus, H. parviplexus, and H. meridionalis in having a ventral sucker less than half the diameter of the oral sucker. It differs from these species by the extension of the vitellaria, which are shorter in the new species, and in the shape of the tegumental spines, which are blunt in the new species. It differs from all other known species of Haematoloechus in the distribution of the uterine loops that are arranged diagonally and that present several short, extracecal. longitudinal loops in the postacetabular region. The new species shows 1.2% sequence divergence in partial 28S sequence with respect to H. medioplexus, 1.1% with H. parviplexus. and 2.5% with H. meridionalis, sequence divergences complementing the morphological differences.     

New genus of Pharyngodonidae (Nematoda: Oxyuridea) and other helminths in Platymantis nexipus (Anura: Ranidae) from Papua New Guinea

Rokroknema novaebritanniae n. gen., n. sp. (Nematoda: Pharyngodonidae) from the large intestine of Platymantis nexipus (Anura: Ranidae) is described and illustrated. Rokroknema represents the second Australo-Papuan genus assigned to the family Pharyngodonidae known to infect frogs. It is similar to Parathelandros (the other genus), but it is readily distinguished because the posterior pair of caudal papillae is not in the form of a rosette. Platymantis nexipus also was found to harbor 1 species of Digenea, Opisthioglyphe cophixali, and 5 additional species of Nematoda, adults of Aplectana krausi, Cosmocerca novaeguineae, Falcaustra batrachiensis, Icosiella papuensis, and larvae in cysts of Abbreviata sp., plus unidentified cystacanths of 1 species of acanthocephalan.