Comparative transcriptome analysis of gonads in male and female Pseudobagrus ussuriensis (Bagridae, Siluriformes).

As an important aquaculture fish in the Heilongjiang River Basin, Pseudobagrus ussuriensis has high economic value, and all-male culture is beneficial to the economic development of this fish. In this study, the transcriptomes of gonads in males and females were analyzed, and some genes related to gonad development were found. A total of 82,931 unigenes were found (average length 1504 bp, N50 1829 bp). In addition, 4689 differentially expressed genes (DEGs; including 1424 genes upregulated and 3265 genes downregulated in males) were identified. Some genes associated with testis development (such as Dmrt1 and Ropn1l) were significantly upregulated in males, while genes related to ovary development (such as Wnt2, PLC, Cyp19a, ZP3) were significantly downregulated in males, demonstrating that these genes have a crucial influence on gonad development in P. ussuriensis. Some signaling pathways related to gonad development were found, such as the Wnt pathway and oocyte meiosis. The results of RNA-seq obtained in this study provide theoretical data for elucidating the potential mechanism of gonad development of P. ussuriensis and reliable genomic data for the establishment of mono-sex breeding of P. ussuriensis.

Concentrations and Risk Assessment of Heavy Metals in Tissues of Walleye Pollock (Gadus chalcogrammus) Captured from the Northeastern Coast of Korea.

HIGHLIGHTS: All heavy metals, except Hg, were well accumulated in liver compared with muscle and gonads. Estimated daily intakes of heavy metals were within 2% of the provisional maximum tolerable daily intakes, Hazard quotients of heavy metals were less than 1.0. Appropriate intake control of G. chalcogrammus is necessary to protect human health in the future.

Unaltered Hypothalamic Metabolic Gene Expression in Kiss1r Knockout Mice Despite Obesity and Reduced Energy Expenditure.

Kisspeptin controls reproduction by stimulating gonadotrophin-releasing hormone neurones via its receptor Kiss1r. Kiss1r is also expressed other brain areas and in peripheral tissues, suggesting additional nonreproductive roles. We recently determined that Kiss1r knockout (KO) mice develop an obese and diabetic phenotype. In the present study, we investigated whether Kiss1r KOs develop this metabolic phenotype as a result of alterations in the expression of metabolic genes involved in the appetite regulating system of the hypothalamus, including neuropeptide Y (Npy) and pro-opiomelanocortin (Pomc), as well as leptin receptor (Lepr), ghrelin receptor (Ghsr), and melanocortin receptors 3 and 4 (Mc3r, Mc4r). Body weights, leptin levels and hypothalamic gene expression were measured in both gonad-intact and gonadectomised (GNX) mice at 8 and 20 weeks of age that had received either normal chow or a high-fat diet. We detected significant increases in Pomc expression in gonad-intact Kiss1r KO mice at 8 and 20 weeks, although there were no alterations in the other metabolic-related genes. However, the Pomc increases appeared to reflect genotype differences in circulating sex steroids, because GNX wild-type and Kiss1r KO mice exhibited similar Pomc levels, along with similar Npy levels. The altered Pomc gene expression in gonad-intact Kiss1r KO mice is consistent with previous reports of reduced food intake in these mice and may serve to increase the anorexigenic drive, perhaps compensating for the obese state. However, the surprising overall lack of changes in any of the hypothalamic metabolic genes in GNX KO mice suggests that the aetiology of obesity in the absence of kisspeptin signalling may reflect peripheral rather than central metabolic impairments.

A timecourse analysis of systemic and gonadal effects of temperature on sexual development of the red-eared slider turtle Trachemys scripta elegans.

Temperature dependent sex determination (TSD) is the process by which the environmental temperature experienced during embryogenesis influences the sex of an organism, as in the red-eared slider turtle Trachemys scripta elegans. In accord with current paradigms of vertebrate sex determination, temperature is believed to exert its effects on sexual development in T. scripta entirely within the middle third of development, when the gonad is forming. However, whether temperature regulates the transcriptome in T. scripta early embryos in a manner that could influence secondary sex characteristics or establish a pro-male or pro-female environment has not been investigated. In addition, apart from a handful of candidate genes, very little is known about potential similarities between the expression cascade during TSD and the genetic cascade that drives mammalian sex determination. Here, we conducted an unbiased transcriptome-wide analysis of the effects of male- and female-promoting temperatures on the turtle embryo prior to gonad formation, and on the gonad during the temperature sensitive period. We found sexually dimorphic expression reflecting differences in steroidogenic enzymes and brain development prior to gonad formation. Within the gonad, we mapped a cascade of differential expression similar to the genetic cascade established in mammals. Using a Hidden Markov Model based clustering approach, we identified groups of genes that show heterochronic shifts between M. musculus and T. scripta. We propose a model in which multiple factors influenced by temperature accumulate during early gonadogenesis, and converge on the antagonistic regulation of aromatase to canalize sex determination near the end of the temperature sensitive window of development.

Thyroid disruption in male goldfish (Carassius auratus) exposed to leachate from a municipal waste treatment plant: Assessment combining chemical analysis and in vivo bioassay.

Several classes of thyroid-disrupting chemicals (TDCs) have been found in refuse leachate, but the potential impacts of leachate on the thyroid cascade of aquatic organisms are yet not known. In this study, we chemically analyzed frequently reported TDCs, as well as conducted a bioassay, to evaluate the potential thyroid-disrupting effects of leachate. We used radioimmunoassay to determine the effects of leachate exposure on plasma 3,3',5-triiodo-l-thyronine (T3), 3,3',5,5'-l-thyroxine (T4), and thyroid-stimulating hormone (TSH) levels in adult male goldfish (Carassius auratus). We also investigated the impacts of leachate treatment on hepatic and gonadal deiodinases [types I (D1), II (D2), and III (D3)] and gonadal thyroid receptor (TRα-1 and TRß) mRNA expressions by using real-time polymerase chain reaction. The results indicated the presence of five TDCs (bisphenol A, 4-t-octylphenol, di-n-butyl phthalate, di-n-octyl phthalate, and diethylhexyl phthalate); their mean concentrations in the leachate were 18.11, 2.76, 4.86, 0.21, and 9.16 µg/L, respectively. Leachate exposure induced plasma T3 and TSH levels in male fish, without influencing the plasma T4 levels. The highly elevated D2 mRNA levels in the liver were speculated to be the primary reason for the induction of plasma T3 levels. Disruption of thyroid functions by leachate was also suggested by the up-regulation of D1 and D2 as well as TRα-1 mRNA levels in the gonads. Prominent thyroid disruptions despite the very low TDC concentrations in the exposure media used in the bioassay strongly indicated the existence of unidentified TDCs in the leachate. Our study indicated the necessity of conducting in vivo bioassays to detect thyroid dysfunctions caused by leachate.

Histopathological baseline levels and confounding factors in common sole (Solea solea) for marine environmental risk assessment.

Liver and gonad histopathology, biometric parameters and hepatic metal bioaccumulation were assessed monthly over a one-year period in common soles from the Basque continental shelf, in order to determine baseline levels and confounding factors within biomonitoring studies. Biometric parameters and hepatic metal bioaccumulation varied according to season and gender. Accordingly, hepatic histopathological traits presented seasonal variations related to the reproductive cycle. However, the hepatic histopathological index showed that seasonality and gender were not significant confounding factors. Conversely, the gonad histopathological index was modulated by season and gender. As for organ comparison, the liver endured more severe histopathological damage than the gonad. In brief, the sampling period and gender may not affect the estimation of hepatic histopathological indices for biomonitoring purposes. Nonetheless, due to different sensitivities to environmental 'noise' variables, the sampling period and gender differentiation should be thoroughly considered for the assessment of gonad histopathology, biometrics and metal bioaccumulation.

Assessment of the health status of wild fish inhabiting a cotton basin heavily impacted by pesticides in Benin (West Africa).

To determine the impact of agricultural pesticides used in cotton cultivation on the health status of fish living in a Beninese cotton basin, we compared the reproductive and hepatic systems of fish sampled from rivers located in both contaminated and pristine conditions. Different types of biomarkers, including biometric indices (a condition factor K, a gonadosomatic index GSI, and a hepatosomatic index HSI), plasma levels of sex steroids (11-ketotestosterone 11-KT, testosterone T and estradiol-17ß E2) and the histopathology of the gonads and liver, were investigated for two different trophic levels of the following two fish species: the Guinean tilapia Tilapia guineensis and the African catfish Clarias gariepinus. The fish were captured during both the rainy season (when there is heavy use of pesticides on cotton fields) and the dry season from one site, in Pendjari River (reference site), which is located outside the cotton-producing basin, and from three other sites on the Alibori River within the cotton-producing basin. Comparing fish that were sampled from contaminated (high levels of endosulfan, heptachlor and DDT and metabolites) and reference sites, the results clearly indicated that agricultural pesticides significantly decreased K and GSI while they increased HSI, regardless of the season, species and sex of the fish. These pesticides also induced a decrease in the plasma levels of 11-KT and T and increased those of E2. The histopathology of the testes revealed, in both species, a high rate of testicular oocytes, up to 50% in the African catfish, downstream of the Alibori River, which indicated estrogenic effects from the pesticides. The disruption of male spermatogenesis primarily included necrosis, fibrosis and the presence of foam cells in the lobular lumen. The histopathology of the ovaries revealed high levels of pre-ovulatory follicular atresia, impaired oogenesis, a decrease in the oocyte vitellogenic diameter and other lesions, such as fibrosis, vacuolation and melano-macrophagic centers. The histopathology of the liver revealed the presence of necrosis, hypertrophic hepatocytes, foci of vacuolation, glycogen depletion and hemosiderin. An assessment of the general health of the fish indicated that all of the sampled fish from the polluted sites were in poorer health compared with those from the reference site but that the African catfish appeared much more affected than the Guinean tilapia, regardless of the sex and season. In conclusion, the overall results indicated that agricultural pesticides significantly impair the endocrine regulation of fish living in the Beninese cotton basin and that this would most likely be one of the causes of the severe damage observed in the liver and gonads and the reduced health condition.

Integrated assessment of runoff from livestock farming operations: Analytical chemistry, in vitro bioassays, and in vivo fish exposures.

Animal waste from livestock farming operations can contain varying levels of natural and synthetic androgens and/or estrogens, which can contaminate surrounding waterways. In the present study, surface stream water was collected from 6 basins containing livestock farming operations. Aqueous concentrations of 12 hormones were determined via chemical analyses. Relative androgenic and estrogenic activity was measured using in vitro cell assays (MDA-kb2 and T47D-Kbluc assays, respectively). In parallel, 48-h static-renewal in vivo exposures were conducted to examine potential endocrine-disrupting effects in fathead minnows. Mature fish were exposed to surface water dilutions (0%, 25%, 50%, and 100%) and 10-ng/L of 17α-ethynylestradiol or 50-ng/L of 17ß-trenbolone as positive controls. Hepatic expression of vitellogenin and estrogen receptor α mRNA, gonadal ex vivo testosterone and 17ß-estradiol production, and plasma vitellogenin concentrations were examined. Potentially estrogenic and androgenic steroids were detected at low nanogram per liter concentrations. In vitro estrogenic activity was detected in all samples, whereas androgenic activity was detected in only 1 sample. In vivo exposures to the surface water had no significant dose-dependent effect on any of the biological endpoints, with the exception of increased male testosterone production in 1 exposure. The present study, which combines analytical chemistry measurements, in vitro bioassays, and in vivo fish exposures, highlights the integrated value and future use of a combination of techniques to obtain a comprehensive characterization of an environmental chemical mixture.

Optimization of effects-assessment of greenside darter (Etheostoma blennioides) exposed to tertiary treated municipal wastewater based on seasonal changes of reproductive endpoints.

The present study describes the seasonal changes in reproductive endpoints of the greenside darter (Etheostoma blennioides) and its implications for environmental monitoring. Fish collections conducted at the appropriate time for the site-specific sentinel fish species can provide a wide variety of population-level information including recruitment, reproduction, and energy storage. The objectives of the present study were to: 1) characterize seasonal changes in reproductive endpoints of the greenside darter (both sexes) to determine the appropriate period for monitoring of this sentinel species; and 2) evaluate the effect of exposure of this sentinel species to tertiary treated municipal effluent at the selected monitoring period. Based on the selected parameters (gonadosomatic index [GSI], liver somatic index [LSI], condition factor, and in vitro gonadal steroid production [testosterone (T) in both sexes; estradiol (E2) in females; and 11-ketotestosterone (11KT) in males]), the present study provides evidence for the value of collecting darters during recrudescence (late fall/early winter) to ensure temporal stability, minimum variability, and stable steroid production capacity. Darters exposed to tertiary treated municipal effluent tended to be larger and heavier relative to reference fish but did not demonstrate any consistent responses in terms of condition or relative liver size. No effect on gonadal development was observed, even though these tertiary-effluent-exposed fish demonstrated a significant reduction in the ability to produce hormones. The present study suggests that although fish exposed to tertiary treated effluent demonstrate no population-level effects, they are still responding at a physiological level. Documentation of the reproductive cycle of sentinel species allows for selection of the most appropriate sampling period to reduce variability and greatly enhances the reliability and interpretation of biological responses.

Assessment of persistent organic pollutants accumulation and lipid peroxidation in two reproductive stages of wild silverside (Odontesthes bonariensis).

Persistent organic pollutants (POPs) in streamwater can sometimes exceed the guidelines values reported for biota and human protection in watersheds with intensive agriculture. Oxidative stress and cytotoxicity are some of the markers of exposure to POPs in fish. Accumulation of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) as well as lipid peroxidation (LPO) was assessed in wild silverside (Odontesthes bonariensis) from maturation and pre-spawning stages sampled in a typical soybean growing area. Pollutants were quantified by gas chromatography with electron capture detection and LPO by the method of thiobarbituric acid reactive substances. Concentrations of POPs were in the following order: OCPs>PCBs>PBDEs in all organs and stages. Liver, gills and gonads had the highest OCP concentrations in both sexes and stages with a predominance of endosulfan in all samples. Matured individuals, sampled after endosulfan application period, showed higher endosulfan concentrations than pre-spawning individuals. The predominance of endosulfan sulfate could be due to direct uptake from diet and water column, as well as to the metabolism of the parent compounds in fish. The prevalence of p,p'-DDE in liver would also reflect both the direct uptake and the metabolic transformation of p,p'-DDT to p,p'-DDE by fish. The highest levels of PBDEs and PCBs were found in gills and brain of both stages of growth. The pattern BDE-47>BDE-100 in all samples corresponds to pentaBDE exposure. In the case of PCBs, penta (#101 and 110) and hexa-CB congeners (#153 and 138) dominated in the maturation stages and tri (#18) and tetra-CB (#44 and 52) in pre-spawning stages, suggesting biotransformation or preferential accumulation of heavier congeners during gonadal development. Differences in LPO levels in ovaries were associated with growth dilution and reproductive stage. Differences in LPO levels in gills were related with pesticide application periods. As a whole, endosulfan, a current-use pesticide, constituted the main pollutant found in wild silverside reflecting the intense agriculture activity in the study area. Moreover endosulfan was positively correlated with LPO.

Development of molecular resources for an intertidal clam, Sinonovacula constricta, using 454 transcriptome sequencing.

BACKGROUND: The razor clam Sinonovacula constricta is a benthic intertidal bivalve species with important commercial value. Despite its economic importance, knowledge of its transcriptome is scarce. Next generation sequencing technologies offer rapid and efficient tools for generating large numbers of sequences, which can be used to characterize the transcriptome, to develop effective molecular markers and to identify genes associated with growth, a key breeding trait. RESULTS: Total RNA was isolated from the mantle, gill, liver, siphon, gonad and muscular foot tissues. High-throughput deep sequencing of S. constricta using 454 pyrosequencing technology yielded 859,313 high-quality reads with an average read length of 489 bp. Clustering and assembly of these reads produced 16,323 contigs and 131,346 singletons with average lengths of 1,376 bp and 458 bp, respectively. Based on transcriptome sequencing, 14,615 sequences had significant matches with known genes encoding 147,669 predicted proteins. Subsequently, previously unknown growth-related genes were identified. A total of 13,563 microsatellites (SSRs) and 13,634 high-confidence single nucleotide polymorphism loci (SNPs) were discovered, of which almost half were validated. CONCLUSION: De novo sequencing of the razor clam S. constricta transcriptome on the 454 GS FLX platform generated a large number of ESTs. Candidate growth factors and a large number of SSRs and SNPs were identified. These results will impact genetic studies of S. constricta.

Neither male gonadal androgens nor female reproductive costs drive development of sexual size dimorphism in lizards.

Sexual size dimorphism (SSD) is an extensively studied phenomenon in animals, including reptiles, but the proximate mechanism of its development is poorly understood. The most pervasive candidates are: (1) androgen-mediated control of growth, i.e. a positive effect of gonadal androgens (testosterone) on male growth in male-larger species, and a negative effect in female-larger species; and (2) sex-specific differences in energy allocation to growth, e.g. sex with larger reproductive costs should result in smaller body size. We tested these hypotheses in adults of the male-larger lizard Paroedura picta by conducting castrations with and without testosterone implants in males and manipulating reproductive status in females. Castration or testosterone replacement had no significant effect on final body length in males. High investment to reproduction had no significant effect on final body length in intact females. Interestingly, ovariectomized females and females with testosterone implants grew to larger body size than intact females. We did not find support for either of the above hypotheses and suggest that previously reported effects of gonadal androgens on growth in male lizards could be a consequence of altered behaviour or social status in manipulated individuals. Exogenous testosterone in females led to decreased size of ovaries; its effect on body size may be caused by interference with normal ovarian function. We suggest that ovarian factors, perhaps estrogens, not reproductive costs, can modify growth in female lizards and may thus contribute to the development of SSD. This hypothesis is largely supported by published results on the effect of testosterone treatment or ovariectomy on body size in female squamates.

Adverse effects in wild fish living downstream from pharmaceutical manufacture discharges.

A set of biochemical and histological responses was measured in wild gudgeon collected upstream and downstream of urban and pharmaceutical manufacture effluents. These individual end-points were associated to fish assemblage characterisation. Responses of biotransformation enzymes, neurotoxicity and endocrine disruption biomarkers revealed contamination of investigated stream by a mixture of pollutants. Fish from sampled sites downstream of the industrial effluent exhibited also strong signs of endocrine disruption including vitellogenin induction, intersex and male-biased sex-ratio. These individual effects were associated to a decrease of density and a lack of sensitive fish species. This evidence supports the hypothesis that pharmaceutical compounds discharged in stream are involved in recorded endocrine disruption effects and fish population disturbances and threaten disappearance of resident fish species. Overall, this study gives argument for the utilisation of an effect-based monitoring approach to assess impacts of pharmaceutical manufacture discharges on wild fish populations.

A whole life cycle assessment on effects of waterborne PBDEs on gene expression profile along the brain-pituitary-gonad axis and in the liver of zebrafish.

Polybrominated diphenyl ethers (PBDEs) are now found ubiquitously in the aquatic environment and biota, and there is a growing concern that PBDEs may disrupt endocrine systems, leading to reproductive impairments of aquatic animals. In our study, zebrafish (Danio rerio) were exposed to the 5 ng/L, 1 µg/L and 50 µg/L of DE-71 for the duration of the whole life cycle (120 days, from eggs to adults). The expression of selected genes along the brain-pituitary-gonadal (BPG) axis and liver, and the levels of plasma sex hormones were examined. In male fish, up-regulation of GnRH in brain, FSHß and LHß in pituitary, FSH-receptor, LH-receptor, and CYP19a in testis was clearly evident, while down-regulation of CYP11a and 3ß-HSD was found in testis. In female fish, a 2.4-fold up-regulation of 3ß-HSD was found in ovary upon exposure to 50 µg/L of DE-71. GnRH in brain, FSHß and LHß in pituitary were also up-regulated, while ERß, TH and TPH in brain and GnRH-receptor in pituitary were significantly down-regulated. Hepatic ERα, AR and VTG in males were all down-regulated, while hepatic ERα and AR in female were up-regulated. Serum estradiol (E2) was reduced in both male and female upon exposure to DE-71, while significant increases in serum testosterone (T) and 11-keto-testosterone (11-KT) were only found in male but not female fish. The ratio of T/E2 as well as the ratio of 11-KT/E2 in male fish increased in a dose-dependent manner upon exposure to DE-71. Our overall results showed that whole life exposure of DE-71 altered the expression of regulatory genes and receptors at all three levels of the BPG axis in zebrafish, and the responses are sex dependent. The observed disruption of GnRH and GtHs can be further related to the subsequent disruption in both levels and balance sex steroid hormones.

Aquatic toxicity assessment of the additive 6-methylcoumarine using four experimental systems.

The toxicity assessment of chemicals is one of the main issues in the current policies in order to protect the health of the environment and human beings. Food and cosmetic additives have been extensively studied in relation to their toxicity to humans, but data about their ecotoxicological effects are scarce. The aim of this study was to evaluate the toxic effects of the additive 6-methylcoumarine in the aquatic milieu using a test battery comprising experimental model systems from different trophic levels. The inhibition of bioluminiscence was studied in the bacteria Vibrio fischeri (decomposer), the inhibition of growth was evaluated in the alga Chlorella vulgaris (producer) and immobilization was studied in the cladoceran Daphnia magna (first consumer). Finally, several end points were evaluated in the RTG-2 salmonid fish cell line, including neutral red uptake, protein content, methylthiazol tetrazolium salt metabolization, glucose-6-phosphate dehydrogenase activity, lactate dehydrogenase activity and leakage, and morphology. The sensitivity of the test systems employed was as follows: V. fischeri > D. magna > C. vulgaris > RTG-2 cell line. The results show that 6-methylcoumarine is not expected to produce acute toxic effects on the aquatic biota. However, chronic and synergistic effects with other chemicals cannot be excluded and should be further investigated.

Karyotype description of Pomacea patula catemacensis (Caenogastropoda, Ampullariidae), with an assessment of the taxonomic status of Pomacea patula

Mitotic chromosomes of the freshwater snail Pomacea patula catemacensis (Baker 1922) were analyzed on gill tissue of specimens from the type locality (Lake Catemaco, Mexico). The diploid number of chromosomes is 2n = 26, including nine metacentric and four submetacentric pairs, therefore, the fundamental number is FN = 52. No sex chromosomes could be identified. The same chromosome number and morphology were already reported for P. flagellata, i.e., the other species of the genus living in Mexico. The basic haploid number for family Ampullariidae was reported to be n = 14 in the literaure; so, its reduction to n = 13 is probably an apomorphy of the Mexican Pomacea snails. Lanistes bolteni, from Egypt, also shows n = 13, but its karyotype is much more asymmetrical, and seems to have evolved independently from P. flagellata and P. patula catemacencis. The nominotypical subspecies, P. patula patula (Reeve 1856), is a poorly known taxon, whose original locality is unknown. A taxonomical account is presented here, and a Mexican origin postulated as the most parsimonious hypothesis.

Reproductive assessment of Japanese medaka (Oryzias latipes) following a four-week fluoxetine (SSRI) exposure.

The occurrence of environmental pharmaceutical products has recently received considerable attention, but impacts on the aquatic environment are largely unknown. Fluoxetine is a widely prescribed antidepressant and acts physiologically as a selective serotonin reuptake inhibitor (SSRI). To determine its potential to disrupt teleost reproductive function, Japanese medaka (Oryzias latipes) were exposed to fluoxetine at aqueous nominal concentrations of 0, 0.1, 0.5, 1 and 5 microg/L for 4 weeks. The last 14 days of this exposure included a reproductive assessment in which no significant changes were observed in egg production, rate of fertilization and spawning, or hatching success of fertilized eggs. A low incidence (1.97-2.53%; 4.02-5.16-fold greater than controls) of developmental abnormalities was observed in offspring from all fluoxetine treatments. Adult gonadal somatic index, hepatic vitellogenin, and ex vivo gonadal steroidogenesis were also unaffected. Circulating plasma estradiol levels in females were significantly increased by 0.1 and 0.5 microg/L treatments. Our study provides novel information on fish biochemical, physiological, and reproduction responses to environmentally realistic fluoxetine concentrations.

Karyotype description of Pomacea patula catemacensis (Caenogastropoda, Ampullariidae), with an assessment of the taxonomic status of Pomacea patula.

Mitotic chromosomes of the freshwater snail Pomacea patula catemacensis (Baker 1922) were analyzed on gill tissue of specimens from the type locality (Lake Catemaco, Mexico). The diploid number of chromosomes is 2n = 26, including nine metacentric and four submetacentric pairs; therefore, the fundamental number is FN = 52, No sex chromosomes could be identified. The same chromosome number and morphology were already reported for P. flagellata, i.e., the other species of the genus living in Mexico. The basic haploid number for family Ampullariidae was reported to be n = 14 in the literature; so, its reduction to n = 13 is probably an apomorphy of the Mexican Pomacea snails. Lanistes bolteni, from Egypt, also shows n = 13, but its karyotype is much more asymmetrical, and seems to have evolved independently from P. flagellata and P. patula catemacensis. The nominotypical subspecies, P. patula patula (Reeve 1856), is a poorly known taxon, whose original locality is unknown. A taxonomical account is presented here, and a Mexican origin postulated as the most parsimonious hypothesis.