Surface ultrastructure of the advanced third-stage larvae of Gnathostoma nipponicum

The surface ultrastructure of advanced third-stage larvae (AL3) of Gnathostoma nipponicum was studied using scanning electron microscopy. The larvae were recovered from the grass snake Rhabdophis tigrina in the Republic of Korea. Parasites had a globular head bulb with a pair of lips at the anterior end and 2 labial papillae and an amphid on each lip. The head bulb was characteristically armed with 3 transverse rows of hooklets, averaging 36, 38, and 43 in number, increasing posteriorly. A total of 213-232 minute unidentate cuticular spines were present along the entire length of the larvae, forming the transverse striations. Two pairs of cervical papillae were located between the 8th and 12th transverse striations, and a pair of body papillae was seen laterally on the posterior third of the body. A pair of caudal phasmids was recognized near the posterior extremity. The surface ultrastructure of AL3 of G. nipponicum is unique compared with that of other species.     

Gnathostoma binucleatum (Spirurida: Gnathostomatidae) from the freshwater fish in Tabasco, Mexico

Human gnathostomiasis is a food-born parasitic disease of relative importance in many countries in Southeast Asia. It is caused by several species of nematodes of the genus Gnathostoma. In Mexico is an emerging public health problem since 1970, when first cases were reported. Until today, larval morphometric characters that have been proposed to differentiate between the three species of Gnathostoma present in this country, are not satisfactory. Recently, the presence of advanced third-stage larvae AdvL3 (infective form for humans) in freshwater fishes from Pantanos de Centla, Tabasco. was recorded but their specific identity was not clarified . Examination of four species of freshwater fishes from the same locality revealed that three of them: Petenia splendida (n=58), Cichlasoma managuense (n=35) and Gobiomorus dormitor (n=9) were infected by 15 AdvL3 of Gnathostoma binucleatum. Specific identity was obtained comparing the internal transcribed spacer 2 (ITS2) of the ribosomal DNA with sequences reported in Genbank. This is the first record of G. binucleatum in P. splendida and G. dormitor from Tabasco and the first specific determination of the parasite in the locality.     

Confirmation of Gnathostoma binucleatum Almeyda-Artigas, 1991, advanced third-stage larvae in Tres Palos Lagoon, Mexico, by morphological and molecular data

Advanced third-stage larvae of Gnathostoma sp. corresponding to 6 morphotypes, distinguished on the basis of the shape of the cephalic hooklets, were collected from the muscle tissue of 5 species of freshwater fish (i.e., Dormitator latifrons, Eleotris picta, Gobiomorus maculatus, Ariopsis guatemalensis, and Cichlasoma trimaculatum) in Tres Palos Lagoon, Guerrero, Mexico. Principal components analysis of 4 morphological characters cluster all samples in a single taxonomic group. A minimum amount of variation was observed among ITS2 sequences of 3 morphotypes and Gnathostoma binucleatum obtained from GenBank (0-0.84%). The observed variation among morphotypes 1, 2, and 3 is the result of intraspecific variability of G. binucleatum supported by morphology and DNA. Morphotypes 4, 5, and 6 belong to the same taxon on the basis of morphology of the hooklets only. For an accurate morphological diagnosis of the causative agent of gnathostomiasis, it is necessary to develop similar studies with other species of the genus.     

Gnathostoma binucleatum: excretion-secretion antigen analysis obtained from advanced third-stage larvae in in vitro culture

The paper describes an introductory characterization of antigenic stimulation of excretion-secretion products (ESP) of Gnathostoma binucleatum advanced third-stage larvae cultured in vitro and proteinases present in this products. Excretory and secretory proteins were obtained after 10 larvae were maintained in 5% CO(2) RPMI medium. The supernatant was collected each week for two months. The proteins were dialyzed, concentrated, and separated in 10% SDS-PAGE gels under reducing conditions and transferred to nitrocellulose paper for immunoblot analyses. G. binucleatum immunized mice serum was used to determine protein antigenicity. Four proteins of 40, 80, 120, and 208 kDa persisted for two months and three proteins, 80, 120, and 208 kDa were recognized for antibodies of mice. In SDS-PAGE gelatin substrate gels ESP resolved as two proteins with molecular weight of 80 and 208 kDa that were sensitive to a metalloproteinase inhibitor, and thus it may be inferred that they might be used for diagnosis of gnathostomiasis.     

Onchocerca volvulus: immunization of chimpanzees with X-irradiated third-stage (L3) larvae.

To provide a theoretical basis for the potential development of vaccines against Onchocerca volvulus (Ov) a trial has been conducted to assess the protective efficacy of immunization of chimpanzees with X-irradiated L3 larvae. Approximately 1000 larvae were injected at 0, 1, and 7 months. The immunized animals, and unimmunized controls, were then challenged with 250 live L3. In order to provide possibly protective exposure to the immunologically distinct L4 epicuticle, a radiation dose (45 krad) was chosen which preserved about 50% of the molting ability of unirradiated larvae. Despite the presence of a strong immune response to crude adult worm extracts, and to cloned Ov antigens, at the time of challenge little or no significant protection against patent infection was observed: three of four immunized animals developed patent infection as compared to four of four controls. One immunized animal failed to become patent or to manifest the late antibody response to adult worm antigens seen in both subpatent and patent infections in this model, and may have been protected from infection. The implications of these studies for future attempts to immunize against O. volvulus are discussed.     

The mosquitoes (Diptera: Culicidae) of Tabasco,Mexico

To document and update the mosquito species of Tabasco, Mexico, field collection trips were conducted in the two physiographic regions of Tabasco: the coastal plain of the southern gulf and the mountains of Chiapas and Guatemala. Mosquitoes were collected as immature and adult stages during the dry and rainy seasons from 2014 through 2015. Additionally, the Reference Collection of Arthropods of Medical Importance (CAIM‐InDRE) containing mosquitoes of Tabasco was re‐examined. In total, 4,913 specimens were collected and examined, which are divided into seven tribes, 18 genera, 27 subgenera, and 104 species. Of these, one genus (Shannoniana Lane and Cerqueira), two subgenera (Georgecraigius Reinert, Harbach and Kitching, and Carrollia Lutz), and 21 species are new records for the mosquito fauna of Tabasco. Culex metempsytus Dyar is a new record for Mexico and Wyeomyia jocosa (Dyar and Knab) is removed from the Mexican mosquito fauna. Seventeen species historically reported were not found in the field collections conducted here. Taxonomic notes, new distribution limits, and comments about the medical importance of species of mosquitoes of Tabasco are discussed. Tabasco is the second state in Mexico with the largest mosquito richness (104 species), followed by Veracruz with 139 species.     

Molecular identification of two strains of third-stage larvae of Contracaecum rudolphii sensu lato (Nematoda: Anisakidae) from fish in Poland

Contracaecum sp. larvae (L3) from fish were identified using nucleotide sequences of the internal transcribed spacers ITS-1 and ITS-2 of the ribosomal DNA. The nematode larvae originated from fish in a freshwater situation (crucian carp Carassius carassius, from Selment Wielki Lake in Mazury, northeastern Poland) and a brackish-water region (Caspian round goby Neogobius melanostomus from the Baltic Sea, Gdafisk Bay at the Polish coast). Two strains (Contracaecum rudolphii A and B) of Contracaecum rudolphii senso lato, a parasite common at the adult stage in fish-eating birds, were identified. In fish from the freshwater site, only the strain temporarily designated C. rudolphii B was identified; in the brackish-water region, both strains were found, suggesting that fish serve as paratenic host for both genotypes. Contracaecum rudolphii sensu lato has been recorded in several species of fish-eating birds in Poland, particularly in the great cormorant, Phalacrocorax carbo, in which the abundance is highest. The results, although based on a restricted number of larvae, suggest that the life cycles of both genotypes can be completed in the Polish region and that at least one of them, C. rudolphii B, can develop both in fresh and brackish water.     

A new species of Ceratozamia (Zamiaceae) from Tabasco and Chiapas, Mexico

Ceratozamia becerrae sp. nov. is described and illustrated. This species from Tabasco and Chiapas has affinity with C. miqueliana H. Wendl. from Veracruz and Chiapas, but differs in morphology and habit of leaves, leaflets, male and female strobili and trunk. Ceratozamia becerrae is considered part of the C. miqueliana species complex that includes C. miqueliana, C. euryphyllidia Vázq.Torres, Sabato & Stevenson and C. zoquorum Pérez-Farrera, Vovides & Iglesias. The geographical range of this species complex is southern Veracruz, Tabasco and northern Chiapas in tropical rain forests.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 123–128.     

Studies on the cryopreservation of Dictyocaulus viviparus (Nematoda) third-stage larvae

Various cooling (0.1-5,100 degrees C min-1) and warming (20-6,800 degrees C min-1) rates, stepped cooling schedules and four cryoprotective additives (dimethyl sulphoxide, methanol, ethanediol and glycerol) were investigated in cryopreservation studies with Dictyocaulus viviparus third-stage larvae. Exsheathment with sodium hypochlorite was essential to achieve significant survival. With uninterrupted cooling, highest survival (30% normally motile) was achieved with rates of 10-70 degrees C min-1. Survival was higher (50-75%) using 1 degree C min-1 to -10 degrees C followed by plunging into liquid nitrogen. The optimum warming rate was 6,800 degrees C min-1. The use of cryoprotectants led to marginally lower survival while varying the suspending media had no significant effect on survival. X-irradiated, exsheathed third-stage larvae cryopreserved by the optimum protocol yielded 38.3 +/- 4.2% survival. Two calves each infected with 45,000 (15,000 viable) exsheathed, unirradiated, cryopreserved third-stage larvae harboured 494 worms (1.1% infectivity) and 355 worms (0.8%) at necropsy. Numbers of first-stage larvae in the faeces reached 420/g and 105/g respectively 27 days after infection.     

Presence and spatio-temporal habitat characterization of Dermatemys mawii (Testudines: Dermatemydidae) in the Grijalva-Usumacinta watershed, Tabasco, Mexico

The Central American River Turtle (Dermatemys mawii) is an endangered species that has been poorly studied. There are no reports on their population status, habitat condition, and the species distribution area is still unknown. This study analyzes the seasonal and spatial variations of their habitat and the presence/absence of D. mawii in three rivers within the Pantanos de Centla Biosphere Reserve (Tabasco, Mexico). For habitat characterization, natural segmentation of rivers was used and three sites per segment were identified, 9 in each rivers (Grijalva and Usumacinta) and 6 in Tabasquillo. Additionally, the evaluation of 11 environmental variables such as water hydrological, physicochemical characteristics and riparian and hydrophytic vegetation were carried out during two different seasons (dry and rainy). The presence/absence of species was assessed with eight fike nets that were set per segment, with a capture effort of 384 hours per trap. The capture per unit effort (CPUE) was used as an indicator of relative abundance. The results indicated spatio-temporal variations in habitat characteristics and the presence of environmental gradients. The principal components analysis (PCA) applied allowed us to determine that the first three components explained 67.8% of the environmental variability. The species presence was confirmed in all rivers, however significant differences exists in their relative abundance: the highest was registered in the Tabasquillo River where the species was present in both seasons and in all segments. Of the 11 environmental variables analyzed, the gradient, shelter and depth were the most indicative of species presence. The obtained results evidenced the importance of riparian vegetation as habitat for Dermatemys. This represents the first approach towards an action plan for a species and its habitat protection within the Pantanos de Centla Biosphere Reserve.     

Diversity of insects captured by weaver spiders (Arachnida: Araneae) in the cocoa agroecosystem in Tabasco, Mexico

The aim of this work was to know the diversity of insects captured by weaver spiders in a plantation of cocoa (Theobroma cacao L.) of 6 ha in the State of Tabasco, Mexico. The study was carried out from July 2004 to June 2005 by means biweekly samples of the insects captured on the spiders webs. The total of 3,041 webs of 54 species of spiders belonging to seven families (Araneidae, Theridiidae, Tetragnathidae, Uloboridae, Pholcidae, Dyctinidae and Linyphiidae) were revised. We found 1,749 specimens belonging to 10 orders of insects, represented by 93 families, the majority of Coleoptera, Diptera and Hemiptera that constituted 74% of the identified families. The biggest number of specimens of all orders was captured by Araneidae, except of Isoptera, whose specimens were captured mainly by the family Theridiidae. The index of diversity (H'), evenness (J') and similarity (Is), applied to know the diversity of families of insects captured among families of spiders, varied from 0.00 to 3.24, 0.00 to 0.81, and 0.04 to 0.522, respectively. We conclude that there is a wide diversity of insects predated by the weaver spiders in the cocoa agroecosystem, and that there are species that can be promising for the biological control of pests.     

Molecular identification of Ammodytes (PISCES, Ammodytidae) larvae, with ontogenetic evidence on separating populations

A total of ten larval Ammodytes were collected from the Yellow (5 specimens) and Bering Seas (5), and a 638 base pair sequence from their mitochondrial DNA COI genes was analyzed for species identification. Sequences were compared with those of adult Ammodytes personatus from the East (4), the Yellow (2) and the East China Seas (2), and Japan (3). Average genetic distance was 0.064 between the Yellow Sea larvae and the Bering Sea larvae, but was 0.0043 between the Yellow Sea larvae and all the adult A. personatus except two individuals from the East Sea. NJ-tree showed that five Yellow Sea larvae were closely clustered with adult A. personatus, except for two individuals. The five Bering Sea larvae were located distantly from adult A. personatus, and are thought to be A. hexapterus. We found morphological differences among two populations of A. personatus larvae and A. hexapterus larvae in morphometric characters and pigmentation. Our results indicated that the East Sea population of A. personatus is closer to A. hexapterus than to the Yellow Sea population of A. personatus, suggesting the East Sea population may be part of separate species.     

Ultrastructural changes in the third-stage, infective larvae of ruminant nematodes treated with sainfoin (Onobrychis viciifolia) extract

Plants rich in condensed tannins are an alternative to chemical anthelmintics to control gastrointestinal nematodes (GINs) in ruminants. Previous functional studies have shown that sainfoin extracts affect the two forms of infective larvae (L3), ensheathed and exsheathed. However, the mechanisms of action remain unknown. The aim of this study was thus to compare ultrastructural changes in ensheathed and exsheathed L3 of two GIN species after in vitro contact with sainfoin extracts using transmission electron microscopy. The main changes identified were an alteration of the hypodermis, the presence of numerous vesicles in the cytoplasm and degeneration and/or death of muscular and intestinal cells. The changes suggested similar and nonspecies-specific lesions in the two nematode species. Comparison of the modifications found in the ensheathed vs. exsheathed L3s revealed different locations of the main cellular changes depending on the larval form. It is hypothesized that these spatial differences in lesions are mainly influenced by the presence of the sheath which favors contact between the active compounds and either the cuticle or the digestive tract. Overall, our observations suggest that the functional changes observed in the biology of GIN L3s after contact with sainfoin extracts are mediated through a direct mode of action, i.e. different interactions between the bioactive plant metabolites and the nematode structure depending on the route of contact.     

Nutritional consequences of the transformation from subsistence to commercial agriculture in Tabasco,Mexico

Recent changes in the area of the Plan Chontalpa in Tabasco have greatly reduced the production of subsistence crops by rural families, resulting in decreased crop diversity and a concomitant increase in the degree of dependence on outside sources of food. Results from a nutrition survey of 149 families demonstrate that dietary diversity, dietary quality, and nutritional status of preschool children are negatively associated with lower crop diversity and increased dependence on purchased foods. Dietary deterioration is illustrated by the negative relationship found between nutritional status and increased sugar consumption. The assumption that a rise in income accompanying the adoption of commercial production will automatically lead to improved nutrition is challenged: income levels were not found to be consistently related to nutritional status. Children of families that have converted to cattle production, despite greater land availability and family incomes, do not have improved nutritional status. In the study area, where wages are low and food prices are very high, the value of a higher degree of self-sufficiency in food is recognized, yet families continue to switch to cash crops due to the environmental, economic, and time constraints imposed by the system of commercial agriculture in which they participate. The solution is not to return to traditional subsistence farming, however, but to determine under what conditions a more progressive form of agricultural change can occur.     

Development of third-stage Physaloptera larvae from Crotalus viridis rafinesque, 1818 in cats with notes on pathology of the larvae in the reptile. (Nematoda, Spiruroidea)

The prairie rattlesnake, Crotalus viridis, was found to be commonly infected with third-stage Physaloptera larvae. A total of 112 larvae were fed to 3 laboratory raised cats. Adult worms recovered 42 and 59 days postinfection were identified as P. rara. Observations were made on the pathology of the larvae in the snake.     

Helminth parasites of the tropical gar, Atractosteus tropicus Gill, from Tabasco, Mexico

A total of 8 helminth species were recorded in an examination of 43 tropical gar, Atractosteus tropicus Gill, collected at the Pantanos de Centla Biosphere Reserve, Tabasco, Mexico. The parasite species included 1 adult trematode, 3 metacercariae, 1 cestode, 1 adult nematode, and 2 nematode larvae. Six of these 8 species were rare, with low prevalence (< 17%) and abundance (< 1.0 helminths per examined fish). The larvae of Contracaecum sp. were the most abundant in the sample, constituting 60% of the total helminths (64% prevalence, 3.8 +/- 5.2 abundance), followed by the cestode Proteocephalus singularis, constituting 18% of the worms (30.5% prevalence, 1.1 +/- 3.0 abundance). Species richness, individual parasite abundance, and diversity were low in the infracommunities. The recording of 3 specialist species in the tropical gar confirms that the helminth fauna of gar has an appreciable degree of specificity. This study indicates the importance of ecological determinants of richness in helminth communities of the tropical gar.     

A new species of Gnathostoma (Nematoda: Gnathostomatidae) in Procyon lotor hernandezii from Mexico

Gnathostoma lamothei n. sp., inhabiting the stomach of Procyon lotor hernandezii Wagler, 1831, in Tlacotalpan, Veracruz State, and Rio Sapo, Oaxaca, Mexico, is described. This new species differs from all other congeners by having the posterior half of the body surface covered by rows of tiny round bosses instead of spines, or lacking ornamentations. Sequences of the ITS2 of the ribosomal DNA of G. lamothei n. sp. are compared with sequences of other species of the genus recorded in Mexico; they show a wide divergence (<50%) with Gnathostoma binucleatum Almeyda-Artigas, 1991, and Gnathostoma turgidum Stossich, 1902, and high similarity with Gnathostoma sp. I sequence (99.2%). On the basis of morphometric traits and sequences, previous records of Gnathostoma sp. I (=Gnathostoma procyonis of Almeyda-Artigas et al., 1994, not Chandler, 1942, and Gnathostoma neoprocyonis nomen nudem) in Mexico are referred to as the new species.     

New host records of the nematode Gnathostoma sp. in Mexico

Gnathostomiasis is an emerging zoonosis in Mexico. However, for most endemic zones, the source of human infection has not been established. During 2000-2003, we investigated 2168 vertebrates (2047 fish, 31 amphibians, 4 reptiles, 19 birds and 67 mammals) from 39 localities distributed in nine states. We registered 7 vertebrate species as new hosts for Gnathostoma, and 22 new locality records for this nematode.