Neotropical sisterhood: new species of Gyrodactylus von Nordmann, 1832 (Platyhelminthes: Monogenea) infecting Rhamdia guatemalensis and Rhamdia laticauda (Siluriformes: Heptapteridae) in Mexico.

We describe two new species of monogenean parasites of the genus Gyrodactylus von Nordmann, 1832 infecting Neotropical catfishes (Siluriformes) in southern Mexico: Gyrodactylus chulini n. sp. from 'chulín', Rhamdia laticauda collected in Oaxaca; and Gyrodactylus juili n. sp. from 'juil', Rhamdia guatemalensis from Veracruz. Morphologically, both new taxa are similar to Gyrodactylus spp. infecting catfishes (Siluriformes) in South America. Sequences of the internal transcribed spacers (ITS1-5.8S-ITS2 rDNA), the D2+D3 domains of the large ribosomal subunit (28S rDNA) and the cytochrome oxidase II (COII) gene were obtained from multiple parasite specimens and analysed using Bayesian inference. Phylogenetic hypotheses using ITS rDNA and COII genes, recovered two new Gyrodactylus species from Rhamdia spp.: G. chulini n. sp.; and Gyrodactylus juili n. sp., which are sister species to Gyrodactylus lilianae, a parasite of Rhamdia quelen in Brazil, and show strong affinity to other gyrodactytlids infecting Neotropical catfishes. This suggests that these new taxa, the first gyrodactylids described from Rhamdia spp. in Mexico, co-migrated to Tropical Middle America with their Neotropical catfish hosts, after the emergence of the Isthmus of Panama.

Development of a light microscopy stain for the sclerites of Gyrodactylus von Nordmann, 1832 (Monogenea) and related genera.

Recent developments in semi-automated identification techniques and the increasing ability to rapidly access digital images and taxonomic descriptions offer to increase the range of individuals capable of performing taxonomic identifications. The present study details methodological approaches undertaken in developing a dedicated stain for the visualisation of monogenean haptoral skeletal elements and reproductive sclerites. The histochemical protocols centre around the use of fluorescent dyes and standard light and laser scanning confocal microscopy to support studies of the functional morphology of these hard structures in small, relatively uncompressed specimens, making these structures more amenable to semi-automated analysis and identification techniques. Staining of the sclerites was achieved using a tissue digestion step to remove the tegument and tissues enclosing the sclerites and then staining them in situ with 40 mM chromothrope 2R (C2R) containing 3 mM phosphotungstic acid (PTA) and 0.5% acetic acid (AA) at room temperature for up to 2 days. Visualisation of the armature of the male copulatory organ of warm water Gyrodactylus species was achieved using 40 mM C2R containing 3 mM PTA for 3 days, whilst cold water species were best stained in 6.4 mM C2R for 1 day without an NaOH pre-treatment. The developed techniques allow for good visualisation of the skeletal elements in a number of monogenean groups and promise to assist the preparation and identification/description of specimens. The 2D/3D digital images of specimens prepared in this manner should provide a useful resource for taxonomists and others needing material to assist specimen identification.

Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea).

Despite routine screening requirements for the notifiable fish pathogen Gyrodactylus salaris, no standard operating procedure exists for its rapid identification and discrimination from other species of Gyrodactylus. This study assessed screening and identification efficiencies under real-world conditions for the most commonly employed identification methodologies: visual, morphometric and molecular analyses. Obtained data were used to design a best-practice processing and decision-making protocol allowing rapid specimen throughput and maximal classification accuracy. True specimen identities were established using a consensus from all three identification methods, coupled with the use of host and location information. The most experienced salmonid gyrodactylid expert correctly identified 95.1% of G. salaris specimens. Statistical methods of classification identified 66.7% of the G. salaris, demonstrating the need for much wider training. Molecular techniques (internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP)/cytochrome c oxidase I (COI) sequencing) conducted in the diagnostic laboratory most experienced in the analysis of gyrodactylid material, identified 100% of the true G. salaris specimens. Taking into account causes of potential specimen loss, the probabilities of a specimen being accurately identified were 95%, 87% and 92% for visual, morphometric and molecular techniques, respectively, and the probabilities of correctly identifying a specimen of G. salaris by each method were 81%, 58% and 92%. Inter-analyst agreement for 189 gyrodactylids assessed by all three methods using Fleiss' Kappa suggested substantial agreement in identification between the methods. During routine surveillance periods when low numbers of specimens are analysed, we recommend that specimens be analysed using the ITS-RFLP approach followed by sequencing of specimens with a "G. salaris-like" (i.e. G. salaris, Gyrodactylus thymalli) banding pattern. During periods of suspected outbreaks, where a high volume of specimens is expected, we recommended that specimens be identified using visual identification, as the fastest processing method, to select "G. salaris-like" specimens, which are subsequently identified by molecular-based techniques.

Life-history characteristics of the large Amazonian migratory catfish Brachyplatystoma rousseauxii in the Iquitos region, Peru.

The main life-history traits of the dorado Brachyplatystoma rousseauxii, a large Amazonian catfish undertaking the largest migration known for a freshwater fish species (from the nursery area in the estuary of the Amazon to the breeding zones in the head waters of the western Amazon basin close to the Andes), were determined from a 5 year sampling of >15,000 specimens in the Peruvian Amazon. The breeding season occurred during the falling and low-water periods, which is hypothesized to be an adaptation to maximize the chances of young stages to reach the estuary. The size at first sexual maturity was slightly larger for females than males, c. 91 and 83 cm standard length (L(S)), respectively. Both males and females reproduce for the first time at >3 years old. The fecundity per spawning event ranged from 481,734 to 1,045,284 oocytes for females weighing 25 and 34 kg, respectively. Seasonal variations of body condition were similar among sexes, but differed between immature specimens that had a higher condition during the low-water period and lower condition during rising waters, and mature individuals that showed the opposite pattern. The growth characteristics were estimated by L(S) frequency analysis. For females, the best fitting models gave a mean birth date in August, during the height of the breeding cycle, with the following von Bertalanffy growth function parameters: L(Sinfinity) = 153.3, K = 0.29 and t(0) =- 0.37 years. For males, the best fitting model gave a mean birth date in July, also during the height of the breeding period, with L(Sinfinity) = 142, K = 0.30 and t(0) =- 0.36 years. At a given age, females were systematically larger than males and the size difference increased with age. The largest females sampled (148 cm L(S)) was 11 years old and the largest male (134 cm L(S)) was 9 years old. The mortality estimates were higher for males total (Z) = 1.34, natural (M) = 0.52 and fishing (F) = 0.82 than for females (Z = 0.98, M = 0.50, F = 0.48). The life-history patterns of B. rousseauxii are discussed in light of the available knowledge about this species and the understanding of its complex life cycle.

Copepods and larvae of nematodes parasitizing (correction of parasiting) the white mullet Mugil curema (Valenciennes, 1836): indicators of anthropogenic impacts in tropical coastal lagoons?

The relationship between parasites and environmental stress were studied in two tropical coastal lagoons of Northwest Mexico: Urias estuary (highly polluted) and Teacapan estuary (slightly polluted). Metazoan parasites were examined in 292 white mullet (Mugil curema) specimens collected bimonthly during a year from both systems. Haliotrema mugilinus, Metamicrocotyla macracantha, Ergasilus sp., Caligus sp., Holobomolochus sp., and Lernaeopodidae were found in gills, while Contracaecum sp. larvae III was found liver, hepatic portal vein and kidneys. Ecological indices were influenced by the slightly higher number of parasitic species in Urias compared to Teacapan, as well as the clear dominance of two species: Ergasilus sp. and Contracaecum sp. in both systems. In fact, Ergasilus sp. showed considerably higher abundance in Urias, possibly indicating that its success was a result of adverse conditions affecting the host, while Contracaecum sp showed higher abundances in Teacapan, suggesting that the environmental conditions occurring in Urias could have produced negative impacts on the nematode's infective potential.