Metals(loids) targeting fish eyes and brain in a contaminated estuary - Uncovering neurosensory (un)susceptibility through bioaccumulation, antioxidant and morphometric profiles.

This study examined the susceptibility of fish (Liza aurata) eyes and brain to metals(loids) contamination under realistic exposure conditions. A multidimensional approach was applied to fish caught at a chronically contaminated site (BAR) and at a reference site of the Tagus estuary (Portugal), which comprised metals(loids) accumulation in eyes and brain together with a battery of enzymatic and non-enzymatic antioxidants, as well as brain morphometry (i.e. cell density). Trace element levels in the blood, gills, liver and kidney allowed interpretations on their preferential pathway(s) to the eyes and brain. Metals(loids) accumulation pointed out the elevated vulnerability of the fish eyes at BAR, probably related with the direct waterborne uptake. Pb uptake in L. aurata eyes could be associated both with water and indirect pathways. At the most contaminated site, metals(loids) were on the basis of pro-oxidant conditions in the ocular tissues, while no indication of toxicity was recorded in the brain. Overall, the results disclosed a differential bioaccumulation among fish organs, suggesting that, in the L. aurata population studied, metal organotropism underlie the lower susceptibility of the brain comparing to the eyes. However, mechanisms remain little understood and further work is needed.

Brain morphometric profiles and their seasonal modulation in fish (Liza aurata) inhabiting a mercury contaminated estuary.

Mercury (Hg) is a potent neurotoxicant known to induce important adverse effects on fish, but a deeper understanding is lacking regarding how environmental exposure affects the brain morphology and neural plasticity of specific brain regions in wild specimens. In this work, it was evaluated the relative volume and cell density of the lateral pallium, hypothalamus, optic tectum and molecular layer of the cerebellum on wild Liza aurata captured in Hg-contaminated (LAR) and non-contaminated (SJ) sites of a coastal system (Ria de Aveiro, Portugal). Given the season-related variations in the environment that fish are naturally exposed, this assessment was performed in the winter and summer. Hg triggered a deficit in cell density of hypothalamus during the winter that could lead to hormonal dysfunctions, while in the summer Hg promoted larger volumes of the optic tectum and cerebellum, indicating the warm period as the most critical for the manifestation of putative changes in visual acuity and motor-dependent tasks. Moreover, in fish from the SJ site, the lateral pallium relative volume and the cell density of the hypothalamus and optic tectum were higher in the winter than in summer. Thus, season-related stimuli strongly influence the size and/or cell density of specific brain regions in the non-contaminated area, pointing out the ability of fish to adapt to environmental and physiological demands. Conversely, fish from the Hg-contaminated site showed a distinct seasonal profile of brain morphology, presenting a larger optic tectum in the summer, as well as a larger molecular layer of the cerebellum with higher cell density. Moreover, Hg exposure impaired the winter-summer variation of the lateral pallium relative size (as observed at SJ). Altogether, seasonal variations in fish neural morphology and physiology should be considered when performing ecotoxicological studies in order to better discriminate the Hg neurotoxicity.

Oxidative stress profiles in brain point out a higher susceptibility of fish to waterborne divalent mercury compared to dietary organic mercury.

This study examines, for the first time, the neurotoxicity of Hg(II) and MeHg in fish (Diplodus sargus) in a time-course comparative perspective and considering realistic exposure levels and routes. Both forms followed an identical time-variation pattern of accumulation in the brain, but dietary MeHg was more efficiently transported to the brain. MeHg was substantially eliminated from the brain in 28days of depuration, which did not occur for Hg(II). Moreover, Hg(II) displayed a high neurotoxicity potential, as unveiled by the poor activation of brain antioxidant defenses and recurrent oxidative damage (as protein oxidation), while the opposite was recorded upon MeHg exposure. These results highlight the need to include Hg(II) in future environmental health assessment plans, preventing an underestimation of the risk for wild fish populations, which has probably been occurring due to the long-standing idea of the higher toxicity of MeHg in comparison with inorganic Hg forms.

Minocycline reduces mechanical allodynia and depressive-like behaviour in type-1 diabetes mellitus in the rat.

A common and devastating complication of diabetes mellitus is painful diabetic neuropathy (PDN) that can be accompanied by emotional disorders such as depression. A few studies have suggested that minocycline that inhibits microglia may attenuate pain hypersensitivity in PDN. Moreover, a recent study reported that minocycline has an acute antidepressive-like effect in diabetic animals. Here we studied whether (i) prolonged minocycline treatment suppresses pain behaviour in PDN, (ii) the minocycline effect varies with submodality of pain, and (iii) the suppression of pain behaviour by prolonged minocycline treatment is associated with antidepressive-like effect. The experiments were performed in streptozotocin-induced rat model of type-1 diabetes. Pain behaviour was evoked by innocuous (monofilaments) and noxious (paw pressure) mechanical stimulation, innocuous cold (acetone drops) and noxious heat (radiant heat). Depression-like behaviour was assessed using forced swimming test. Minocycline treatment (daily 80mg/kg per os) of three-week duration started four weeks after induction of diabetes. Diabetes induced mechanical allodynia and hyperalgesia, cold allodynia, heat hypoalgesia, and depression-like behaviour. Minocycline treatment significantly attenuated mechanical allodynia and depression-like behaviour, while it failed to produce significant changes in mechanical hyperalgesia, cold allodynia or heat hypoalgesia. The results indicate that prolonged per oral treatment with minocycline has a sustained mechanical antiallodynic and antidepressive-like effect in PDN. These results support the proposal that minocycline might provide a treatment option for attenuating sensory and comorbid emotional symptoms in chronic PDN.

Unveiling the neurotoxicity of methylmercury in fish (Diplodus sargus) through a regional morphometric analysis of brain and swimming behavior assessment.

The current study aims to shed light on the neurotoxicity of MeHg in fish (white seabream - Diplodus sargus) by the combined assessment of: (i) MeHg toxicokinetics in the brain, (ii) brain morphometry (volume and number of neurons plus glial cells in specific brain regions) and (iii) fish swimming behavior (endpoints associated with the motor performance and the fear/anxiety-like status). Fish were surveyed for all the components after 7 (E7) and 14 (E14) days of dietary exposure to MeHg (8.7µgg-1), as well as after a post-exposure period of 28days (PE28). MeHg was accumulated in the brain of D. sargus after a short time (E7) and reached a maximum at the end of the exposure period (E14), suggesting an efficient transport of this toxicant into fish brain. Divalent inorganic Hg was also detected in fish brain along the experiment (indicating demethylation reactions), although levels were 100-200 times lower than MeHg, which pinpoints the organic counterpart as the great liable for the recorded effects. In this regard, a decreased number of cells in medial pallium and optic tectum, as well as an increased hypothalamic volume, occurred at E7. Such morphometric alterations were followed by an impairment of fish motor condition as evidenced by a decrease in the total swimming time, while the fear/anxiety-like status was not altered. Moreover, at E14 fish swam a greater distance, although no morphometric alterations were found in any of the brain areas, probably due to compensatory mechanisms. Additionally, although MeHg decreased almost two-fold in the brain during post-exposure, the levels were still high and led to a loss of cells in the optic tectum at PE28. This is an interesting result that highlights the optic tectum as particularly vulnerable to MeHg exposure in fish. Despite the morphometric alterations reported in the optic tectum at PE28, no significant changes were found in fish behavior. Globally, the effects of MeHg followed a multiphasic profile, where homeostatic mechanisms prevented circumstantially morphometric alterations in the brain and behavioral shifts. Although it has become clear the complexity of matching brain morphometric changes and behavioral shifts, motor-related alterations induced by MeHg seem to depend on a combination of disruptions in different brain regions.

Inorganic mercury accumulation in brain following waterborne exposure elicits a deficit on the number of brain cells and impairs swimming behavior in fish (white seabream-Diplodus sargus).

The current study contributes to fill the knowledge gap on the neurotoxicity of inorganic mercury (iHg) in fish through the implementation of a combined evaluation of brain morphometric alterations (volume and total number of neurons plus glial cells in specific regions of the brain) and swimming behavior (endpoints related with the motor activity and mood/anxiety-like status). White seabream (Diplodus sargus) was exposed to realistic levels of iHg in water (2µgL(-1)) during 7 (E7) and 14 days (E14). After that, fish were allowed to recover for 28 days (PE28) in order to evaluate brain regeneration and reversibility of behavioral syndromes. A significant reduction in the number of cells in hypothalamus, optic tectum and cerebellum was found at E7, accompanied by relevant changes on swimming behavior. Moreover, the decrease in the number of neurons and glia in the molecular layer of the cerebellum was followed by a contraction of its volume. This is the first time that a deficit on the number of cells is reported in fish brain after iHg exposure. Interestingly, a recovery of hypothalamus and cerebellum occurred at E14, as evidenced by the identical number of cells found in exposed and control fish, and volume of cerebellum, which might be associated with an adaptive phenomenon. After 28 days post-exposure, the optic tectum continued to show a decrease in the number of cells, pointing out a higher vulnerability of this region. These morphometric alterations coincided with numerous changes on swimming behavior, related both with fish motor function and mood/anxiety-like status. Overall, current data pointed out the iHg potential to induce brain morphometric alterations, emphasizing a long-lasting neurobehavioral hazard.

A role of supraspinal galanin in behavioural hyperalgesia in the rat.

INTRODUCTION: In chronic pain disorders, galanin (GAL) is able to either facilitate or inhibit nociception in the spinal cord but the contribution of supraspinal galanin to pain signalling is mostly unknown. The dorsomedial nucleus of the hypothalamus (DMH) is rich in galanin receptors (GALR) and is involved in behavioural hyperalgesia. In this study, we evaluated the contribution of supraspinal GAL to behavioural hyperalgesia in experimental monoarthritis. METHODS: In Wistar-Han males with a four week kaolin/carrageenan-induced monoarthritis (ARTH), paw-withdrawal latency (PWL) was assessed before and after DMH administration of exogenous GAL, a non-specific GALR antagonist (M40), a specific GALR1 agonist (M617) and a specific GALR2 antagonist (M871). Additionally, the analysis of c-Fos expression after GAL injection in the DMH was used to investigate the potential involvement of brainstem pain control centres. Finally, electrophysiological recordings were performed to evaluate whether pronociceptive On- or antinociceptive Off-like cells in the rostral ventromedial medulla (RVM) relay the effect of GAL. RESULTS: Exogenous GAL in the DMH decreased PWL in ARTH and SHAM animals, an effect that was mimicked by a GALR1 agonist (M617). In SHAM animals, an unselective GALR antagonist (M40) increased PWL, while a GALR2 antagonist (M871) decreased PWL. M40 or M871 failed to influence PWL in ARTH animals. Exogenous GAL increased c-Fos expression in the RVM and dorsal raphe nucleus (DRN), with effects being more prominent in SHAM than ARTH animals. Exogenous GAL failed to influence activity of RVM On- or Off-like cells of SHAM and ARTH animals. CONCLUSIONS: Overall, exogenous GAL in the DMH had a pronociceptive effect that is mediated by GALR1 in healthy and arthritic animals and is associated with alterations of c-Fos expression in RVM and DRN that are serotonergic brainstem nuclei known to be involved in the regulation of pain.

Prevalence, intensity, and abundance of infection and pathogenesis caused by diphyllobothriosis in vulnerable, native fish and introduced trout in Lake Panguipulli, Chile.

Diphyllobothriosis in fish from freshwater ecosystems in southern Chile was first reported in 1949. Infection by plerocercoids of Diphyllobothrium latum and Diphyllobothrium dendriticum occurs in introduced trout (Oncorhynchus mykiss) and native fish. We determined the prevalence, mean intensity, and mean abundance of seasonal infection and tissue damage produced by Diphyllobothrium spp. in native fish (Percichthys trucha, Odontesthes mauleanum, and Basilichthys australis) and introduced trout (O. mykiss) from Lake Panguipulli, Chile. Prevalence, mean intensity, and mean abundance of D. latum infection were significantly greater in trout than they were in native fish. Prevalence and mean abundance were similar in O. mauleanum and P. trucha, but they were greater than those in B. australis. Prevalence and abundance were similar among seasons between sexes for the four hosts. For all host species, except P. trucha, there was a statistically significant positive correlation between host length and the abundance of plerocercoids. Infections in muscle tissue were present in 61% of trout compared with 23% in O. mauleanum and 12% in P. trucha, suggesting a greater risk for human infection when consuming trout. In general, prevalence of infection by D. dendriticum was lower than was D. latum prevalence. Encapsulation of plerocercoids was common and severe in 71% of the trout examined. Only slight encapsulation of plerocercoids was found in the native O. mauleanum, and no encapsulation was observed in P. trucha or B. australis. The greater concentration of plerocercoids in the walls of the digestive tract of trout suggests a more-rapid immune response in trout than in native fish. The low frequency of encapsulation of plerocercoids in native fish would mean greater tissue damage in the natives than that observed in the trout because they are free to migrate among the viscera, potentially endangering these native fish populations in regions where Diphyllobothrium spp. are endemic.

Infección por helmintos parásitos en salmón coho, oncorhynchus kisutch, durante su retorno al río Simpson, Chile / Infection by parasitic helmiths in coho sallmon, oncorhynchus kisutch, during their return to Simpson river, Chile

Between may and july 1994, 17 adult returning salmons, oncorhynchus kisutch, were collected in the River Simpson, Chile. All fishes showed infection by plerocercoids of diphyllobothrium sp. in different locations: stomach, spleen, liver, mesenteries and gonads. Infection with larval cestodes of an unidentified species of phillobothriidae was determined in the intestine of seven (41,2 percent) salmons and its prevalences of infection showed significant differences between female and male salmons. The 94,4 percent of total plerocercoids of diphyllobothrium were isolated from the stomach wall. Prevalence and mean intensity of infection by diphyllobothrium sp. did not show significant differences between fishes of different sex

Aplectana artigasi n.sp. (Nematoda: Cosmocercidae) from the frog Eupsophus calcaratus (Anura: Leptodactylidae) in Southern Chile

A new species found in the intestine of Eupsophus calcaratus (Anura: Leptodactylidae) from Yaldad, Chile (45º5'S;73º43'W) is described. This nematode is compared with other South American species of the genus. Aplectana artigasi differs from the only known Chilean species, Aplectana chilensis, as the former has a greater number of postanal papillae and double papilla in the ventral region of the tail.

Nuevo huésped y notas taxonómicas para baerietta chilensis: cestoda nematotaeniidae / A new host and taxonomic notes for baerietta chilensis: cestoda nematotaeniidae

The chilean leptodactylid frog eupsophus calcaratus, a typical inhabitant of the temperate forest of South America, is registered as a new host of baerietta chilensis (cestoda: nematotaeniidae)

Fauna helmintológica en anuros chilenos / Helminthological fauna of chilean anura

In this paper a list of the endoparasites described for the chilean frogs and toads is presented. Also it is indicated the locality of the hosts, prevalence and intensity of the infection. Nine species of parasites have been described; three trematodes: Gorgoderina chilensis in Rhinoderma darwinii, Gorgoderina valdiviensis in Caudiverbera caudiverbera and Rudolphitrema chilensis in Eusophus roseus; two cestodes: Ophiotaenia noei in C. caudiverbera and Baerietta chilensis in Batrachyla taeniata; three nematodes: Aplectana chilensis and Cosmicerca chilensis in R. darwinii and Oswaldocruzia neghmei in Hylorina sylvatica, Eusophus miqueli, Eusophus vertebralis and E. roseus and one acanthocephala: Acanthocephalus caspanensis in Bufo spinulosus

Lepocreadium valdiviensis nov. sp. (digenea: lepocreadiidae) parásito de paralichthys microps lenguado de ojos chicos en el sur de Chile / Lepocreadium valdiviensis nov. sp., (digenea: lepocreadiidae) parasite of paralichthys microps small eyes flounder in south Chile

Se describe una nueva especie de lepocreadium, L. valdiviensis (Digenea) del intestino delgado del lenguado, Paralichthys microps Günther, 1881. difiere de las otras especies del género, principalmente por la distribución de los folículos vitelógenos, los cuales se extienden desde el extremo posterior del cuerpo hasta la ventosa ventral; la porción anterior del cuerpo cubierta con espinas; útero preovárico; ovario y testículos compactos. Este hallazgo extiende la distribución geográfica del género e identifica un nuevo huésped para este taxón