The Virus-Induced Cytopathic Effect.

The cytopathic effect comprises the set of cellular alterations produced by a viral infection. It is of great relevance since it constitutes a direct marker of infection. Likewise, these alterations are often virus-specific which makes them a phenotypic marker for many viral species. All these characteristics have been used to complement the study of the dynamics of virus-cell interactions through the kinetic study of the progression of damage produced by the infection. Various approaches have been used to monitor the cytopathic effect, ranging from light microscopy, immunofluorescence assays, and direct labeling with fluorescent dyes, to plaque assay for the characterization of the infection over time. Here we address the relevance of the study of cytopathic effect and describe different experimental alternatives for its application.

Dosimetry in cranial photobiomodulation therapy: effect of cranial thickness and bone density.

This research aims to examine the influence of human skull bone thickness and density on light penetration in PBM therapy across different wavelengths, focusing on how these bone characteristics affect the absorption of therapeutic light. Analyses explored the effect of skull bone density and thickness on light penetration in PBM, specifically using Low-Level Laser Therapy (LLLT) for efficacy prediction. Measurements of bone thickness and density were taken using precise tools. This approach emphasizes LLLT's significance in enhancing PBM outcomes by assessing how bone characteristics influence light penetration. The study revealed no significant correlation between skull bone density and thickness and light penetration capability in photobiomodulation (PBM) therapy, challenging initial expectations. Wavelengths of 405 nm and 665 nm showed stronger correlations with bone density, suggesting a significant yet weak impact. Conversely, wavelengths of 532 nm, 785 nm, 810 nm, 830 nm, 980 nm, and 1064 nm showed low correlations, indicating minimal impact from bone density variations. However, data variability (R2 < 0.4) suggests that neither density nor thickness robustly predicts light power traversing the bone, indicating penetration capability might be more influenced by bone thickness at certain wavelengths. The study finds that the effectiveness of photobiomodulation (PBM) therapy with bone isn't just based on bone density and thickness but involves a complex interplay of factors. These include the bone's chemical and mineral composition, light's wavelength and energy dose, treatment duration and frequency, and the precise location where light is applied on the skull.

Photobiomodulation increases brain metabolic activity through a combination of 810 and 660 wavelengths: a comparative study in male and female rats.

Photobiomodulation (PBM), an emerging and non-invasive intervention, has been shown to benefit the nervous system by modifying the mitochondrial cytochrome c-oxidase (CCO) enzyme, which has red (620-680 nm) or infrared (760-825 nm) spectral absorption peaks. The effect of a single 810-nm wavelength with a combination of 810 nm and 660 nm lights in the brain metabolic activity of male and female rats was compared. PBM, with a wavelength of 810 nm and a combination of 810 nm and 660 nm, was applied for 5 days on the prefrontal cortex. Then, brain metabolic activity in the prefrontal area, hippocampus, retrosplenial, and parietal cortex was explored. Sex differences were found in cortical and subcortical regions, indicating higher male brain oxidative metabolism, regardless of treatment. CCO activity in the cingulate and prelimbic area, dentate gyrus, retrosplenial and parietal cortex was enhanced in both treatments (810 + 660 nm and 810 nm). Moreover, using the combination of waves, CCO increased in the infralimbic area, and in CA1 and CA3 of the hippocampus. Thus, employment of a single NIR treatment or a combination of red to NIR treatment led to slight differences in CCO activity across the limbic system, suggesting that a combination of lights of the spectrum may be relevant.

Increased basolateral amygdala metabolic activity during flavor familiarization: an experimental study.

BACKGROUND: Novel flavors elicit a cautious neophobic response which is attenuated as the flavor becomes familiar and safe. The attenuation of neophobia reveals the formation of a safe memory. Previous lesion studies in rats have reported that basolateral amygdala integrity is required for taste neophobia, but not neophobia to flavor, i.e., taste linked to an odorous component. Accordingly, immunohistochemical analyses show that novel tastes induced higher basolateral amygdala activity when compared to familiar ones. However, a different role of basolateral amygdala in flavor attenuation of neophobia is suggested by lesion studies using a vinegar solution. Studies assessing basolateral amygdala activity during flavor attenuation of neophobia are lacking. Thus, we quantified cytochrome oxidase as an index of basolateral amygdala activity along the first and second vinegar exposures in order to assess flavor neophobia and attenuation of neophobia. METHODS: We exposed adult male Wistar rats either once or twice to a 3% cider vinegar solution or water, and compared the basolateral amygdala, piriform cortex and caudate putamen brain metabolic activity using cytochrome c-oxidase histochemistry. RESULTS: We found increased flavor intake and cytochrome c-oxidase histochemistry activity during the second exposure in basolateral amygdala, but not in the piriform cortex and caudate/putamen. CONCLUSIONS: The main finding of the study is that BLA metabolic activity was higher in the group exposed to a familiar vinegar solution than in the groups exposed to either water or a novel vinegar solution.

Are there sex differences in spatial reference memory in the Morris water maze? A large-sample experimental study.

Sex differences have been found in allocentric spatial learning and memory tasks, with the literature indicating that males outperform females, although this issue is still controversial. This study aimed to explore the behavior of male and female rats during the habituation and learning of a spatial memory task performed in the Morris Water Maze (MWM). The study included a large sample of 89 males and 85 females. We found that females searched slightly faster than males during habituation with a visible platform. During learning, both male and female rats decreased the latency and distance traveled to find the hidden platform over the days, with males outperforming females in the distance traveled. Females swam faster but did not find the platform earlier, suggesting a less directed navigational strategy. Both sexes increased time spent in the target zone over the days, with no sex differences. Although females swam more in the periphery during the first days of the task, both sexes decreased the time spent in this area. Finally, only males increased swimming in the pool's center over the days, spending more time than females in this area across the entire training. In conclusion, we need to register several variables in the MWM and analyze path strategies to obtain more robust results concerning sex differences. Research on spatial learning should include both sexes to achieve a more equitable, representative, and translational science.

Mental health issues and psychological risk factors among Paraguayan healthcare workers during the COVID-19 pandemic.

BACKGROUND: There is an increasing concern regarding the mental health of healthcare workers during the COVID-19 pandemic. AIMS: This study aimed to assess the psychological impact of the COVID-19 exposure among Paraguayan healthcare workers. METHODS: A cross-sectional descriptive study has been carried out in five hospitals of Paraguay. Demographic and occupational exposure to COVID-19 were collected through a short questionnaire. Mental health status was assessed with the GAD-7, the PHQ-9, and the PCL-C. Logistic regression was used to determine psychological risk factors. RESULTS: 432 participants were surveyed. 218 (50.46%) were physicians. The prevalence of symptoms of anxiety, depression and PTSD was 48.15, 41.90, and 5.79%, respectively. There were no significant differences in anxiety (128 [29.63%] vs. 80 [18.52%]; p = 0.3303), depression (102 [23.61%] vs. 79 [18.29%]; p = 0.6703), or PTSD (14 [3.24%] vs. 11 [2.55%]; p = 0.8074) between frontline versus second-line workers. Main risk factors associated with psychological distress included work experience <5 years for depression and a COVID-19 positive diagnosis or having family/friends with a COVID-19 positive diagnosis for PTSD. CONCLUSIONS: Paraguayan healthcare workers reported high prevalence of anxiety, depression, and a low prevalence of PTSD. A positive diagnosis of COVID-19 and work experience <5 years are important psychological risk factors.

Effect of Adipose-Derived Mesenchymal Stem Cells (ADMSCs) Application in Achilles-Tendon Injury in an Animal Model.

Background: Achilles-tendon rupture prevails as a common tendon pathology. Adipose-derived mesenchymal stem cells (ADMSCs) are multipotent stem cells derived from adipose tissue with attractive regeneration properties; thus, their application in tendinopathies could be beneficial. Methods: Male rabbit ADMSCs were obtained from the falciform ligament according to previously established methods. After tenotomy and suture of the Achilles tendon, 1 × 106 flow-cytometry-characterized male ADMSCs were injected in four female New Zealand white rabbits in the experimental group (ADMSC group), whereas four rabbits were left untreated (lesion group). Confirmation of ADMSC presence in the injured site after 12 weeks was performed with quantitative sex-determining region Y (SRY)-gene RT-PCR. At Week 12, histochemical analysis was performed to evaluate tissue regeneration along with quantitative RT-PCR of collagen I and collagen III mRNA. Results: Presence of male ADMSCs was confirmed at Week 12. No statistically significant differences were found in the histochemical analysis; however, statistically significant differences between ADMSC and lesion group expression of collagen I and collagen III were evidenced, with 36.6% and 24.1% GAPDH-normalized mean expression, respectively, for collagen I (p < 0.05) and 26.3% and 11.9% GAPDH-normalized mean expression, respectively, for collagen III (p < 0.05). The expression ratio between the ADMSC and lesion group was 1.5 and 2.2 for collagen I and collagen III, respectively. Conclusion: Our results make an important contribution to the understanding and effect of ADMSCs in Achilles-tendon rupture.

Hippocampal neuropeptide Y2 receptor blockade improves spatial memory retrieval and modulates limbic brain metabolism.

INTRODUCTION: The neuropeptide Y (NPY) is broadly distributed in the central nervous system (CNS), and it has been related to neuroprotective functions. NPY seems to be an important component to counteract brain damage and cognitive impairment mediated by drugs of abuse and neurodegenerative diseases, and both NPY and its Y2 receptor (Y2R) are highly expressed in the hippocampus, critical for learning and memory. We have recently demonstrated its influence on cognitive functions; however, the specific mechanism and involved brain regions where NPY modulates spatial memory by acting on Y2R remain unclear. METHODS: Here, we examined the involvement of the hippocampal NPY Y2R in spatial memory and associated changes in brain metabolism by bilateral administration of the selective antagonist BIIE0246 into the rat dorsal hippocampus. To further evaluate the relationship between memory functions and neuronal activity, we analysed the regional expression of the mitochondrial enzyme cytochrome c oxidase (CCO) as an index of oxidative metabolic capacity in limbic and non-limbic brain regions. RESULTS: The acute blockade of NPY Y2R significantly improved spatial memory recall in rats trained in the Morris water maze that matched metabolic activity changes in spatial memory processing regions. Specifically, CCO activity changes were found in the dentate gyrus of the dorsal hippocampus and CA1 subfield of the ventral hippocampus, the infralimbic region of the PFC and the mammillary bodies. CONCLUSIONS: These findings suggest that the NPY hippocampal system, through its Y2R receptor, influences spatial memory recall (retrieval) and exerts control over patterns of brain activation that are relevant for associative learning, probably mediated by Y2R modulation of long-term potentiation and long-term depression.

A fluorescence-activatable reporter of flavivirus NS2B-NS3 protease activity enables live imaging of infection in single cells and viral plaques.

The genus Flavivirus in the family Flaviviridae comprises many medically important viruses, such as dengue virus (DENV), Zika virus (ZIKV), and yellow fever virus. The quest for therapeutic targets to combat flavivirus infections requires a better understanding of the kinetics of virus-host interactions during infections with native viral strains. However, this is precluded by limitations of current cell-based systems for monitoring flavivirus infection in living cells. In the present study, we report the construction of fluorescence-activatable sensors to detect the activities of flavivirus NS2B-NS3 serine proteases in living cells. The system consists of GFP-based reporters that become fluorescent upon cleavage by recombinant DENV-2/ZIKV proteases in vitro A version of this sensor containing the flavivirus internal NS3 cleavage site linker reported the highest fluorescence activation in stably transduced mammalian cells upon DENV-2/ZIKV infection. Moreover, the onset of fluorescence correlated with viral protease activity. A far-red version of this flavivirus sensor had the best signal-to-noise ratio in a fluorescent Dulbecco's plaque assay, leading to the construction of a multireporter platform combining the flavivirus sensor with reporter dyes for detection of chromatin condensation and cell death, enabling studies of viral plaque formation with single-cell resolution. Finally, the application of this platform enabled the study of cell-population kinetics of infection and cell death by DENV-2, ZIKV, and yellow fever virus. We anticipate that future studies of viral infection kinetics with this reporter system will enable basic investigations of virus-host interactions and facilitate future applications in antiviral drug research to manage flavivirus infections.

Early life stress due to repeated maternal separation alters the working memory acquisition brain functional network.

Unfortunately, adverse environments in early life are frequently found in most human populations. Early life stress leads to diverse cognitive impairments, some of them related to learning and memory and executive functions such as working memory (WM). We employ an animal model of early stress using repeated maternal separation (MS) for 4 h a day on 21 consecutive days, pre-weaning. In adulthood, we tested their spatial WM using the Morris water maze. MS subjects showed a marked delay in the acquisition of the task. In addition, we explored brain energy oxidative metabolism and found an increase in cytochrome c oxidase (CCO) activity in the cingulate cortex, anterior thalamus, and supramammillary areas, indicating an intense effort to successfully solve the WM task. However, decreased CCO activity was found in the medial-medial mammillary nucleus in MS animals, which would partially explain the delayed acquisition of the WM task. Further studies are needed to explore the long-term alterations produced by early stress. LAY SUMMARY A stressful environment caused by the separation of baby rats from the mother for several hours a day in the first stages of postnatal life can be devastating to brain cells, making them look for alternative sources of energy, among other changes. These alterations in brain functional networks would lead to cognitive impairments such as the delayed acquisition of new learning and strategies.

Retrieval of allocentric spatial memories is preserved up to thirty days and does not require higher brain metabolic demands.

Spatial orientation is a cognitive ability that is indispensable for survival. Several visual distal cues present in the context can be integrated, establishing a cognitive map. Although there is cumulative evidence about the neural substrate involved in spatial memory acquisition, the brain networks mediating the processes involved in the retrieval of allocentric spatial memories have been studied less. Here, we aimed to explore the role of neuronal oxidative metabolism in the retrieval of allocentric spatial memories through cytochrome c oxidase (CCO) histochemistry seven, 15, 30, 45, and 60 days after task acquisition. Our behavioural results show that spatial memory retrieval in male and female rats is preserved seven, 15, and 30 days post-acquisition, but there is forgetfulness after this time, with subjects not being able to remember the position of the hidden platform after 45 and 60 dfearays. Regarding the study of male brain metabolism, we observed reduced CCO activity in the medial prefrontal cortex, the parietal, retrosplenial, rhinal cortex, and the hippocampal regions in all the groups that failed to solve the task. Similar results were found for female brain oxidative metabolism, in addition to certain differences between succefearssful-retrieval female groups. In conclusion, our work adds information about the behavioural retrieval of an allocentric spatial reference task, suggesting that recovering spatial information seven, 15, and 30days after acquisition is a simple task that does not require a high metabolic demand, in both male and female rats.

How Does Maternal Separation Affect the Cerebellum? Assessment of the Oxidative Metabolic Activity and Expression of the c-Fos Protein in Male and Female Rats.

Early life stress increases the risk of abnormal brain development, and it is associated with psychological disorders. Maternal separation is an established animal model of early life stress that produces changes in the development of the central nervous system. The objective of this study was to evaluate the effect of maternal separation on the rat cerebellum, both behaviourally and physiologically. We used 32 rats, males (n = 8) and females (n = 7), subjected to maternal separation for 21 days and a control group (9 males and 8 females). Spatial reference memory was assessed using the Morris water maze, and brain metabolic activity and the expression of an immediate early gene were determined, respectively, using the histochemical technique of cytochrome c oxidase and the immunocytochemistry of c-Fos. Results showed that both groups successfully performed the spatial memory task. Although there were no behavioural differences, the experimental group showed lower metabolic activity in the medial nucleus of the cerebellum, as well as fewer c-Fos-positive cells in the three deep nuclei of the cerebellum. These decreases could contribute to the emotional or behavioural impairments reported in maternal separation subjects.

[Identification of behavioural and emotional problems in premature children in the primary care setting]. / Identificación de problemas de conducta y emocionales en niños prematuros en el ámbito de atención primaria.

OBJECTIVE: To demonstrate the usefulness, for use by the primary care pediatrician, of the BASC questionnaire (Behavior Assessment System for Children) for the early detection of psychological and behavioral problems in premature infants. DESIGN: Cross-sectional and descriptive study. SETTING: Primary care (Health Area IV of the Principado of Asturias) and Hospital Universitario Central de Asturias, Spain. PARTICIPANTS: Parents of 87 premature children with birth weight less than 1500g and 43 full-term controls, both aged 5-7 years. MAIN MEASUREMENTS: The BASC questionnaire (parent version) was applied. RESULTS: Preterm children presented a high rate of inactivity (Z = -4.125, P < 0.001), anxiety (Z = -2.801, P = 0.005) and internalization problems (Z = -2.148, P = 0.032), being more evident at 5 years of age. Preterm boys show higher levels of hyperactivity (Z = -2.082, P = 0.037) and behavioural problems (Z = -2.354, P = 0.019) than girls, who presented more attentional problems (Z = -2.345; P = 0.019). CONCLUSIONS: The BASC questionnaire is useful for the detection and early diagnosis at the primary care level of the behavioral and emotional problems of premature children.

Early life stress by repeated maternal separation induces long-term neuroinflammatory response in glial cells of male rats.

Childhood maltreatment and neglect lead to a wide range of mental disorders highlighted by hormone and immune alterations in neglected children. This social-health challenge has led to the creation of early stress models such as maternal separation (MS) in rodents. We performed a MS model (4 h per day, 21 days; n = 16 MS and n = 16 control), and then measured three parameters in adult male rat brains, in order to look for long-term effects of early life stress. We used immunocytochemistry to mark glial fibrillary acidic protein (GFAP)-positive cells, which indicates changes in astroglia, and ionized calcium binding adaptor molecule 1 (Iba-1)-positive cells, which inform about reactive microglia. In order to study mRNA levels of some immune mediators, interleukin determination (interleukin-6, IL-6; tumor necrosis factor, TNFα) mRNAs were evaluated by real-time polymerase chain reaction (rt-PCR) in discrete brain regions. Measurements of numbers of GFAP-positive cells, and expression of Iba-1, IL-6 and TNFα mRNAs were performed in prefrontal cortex (PFC): cingulate cortex (CG), prelimbic cortex (PL) and infralimbic cortex (IL), striatal areas (dorsal striatum, STD; and nucleus accumbens, ACC), and dorsal hippocampus (HC: CA1, CA3 and dentate gyrus (DG)). We found that MS produces a dramatic and sustained decrease in the astroglial population in all the areas measured (from -25% in CA1 to -85.7% in ACC), whereas increased numbers of microglia were found, in more restricted regions: STD (72.6%), ACC (31%) and CA3 (33.3%) areas. Regarding mRNA measurements, we found increased IL-6 mRNA expression in HC (104.2%), and after MS.

Hepatic encephalopathy: Sometimes more portal than hepatic.

Hepatic encephalopathy is a severe complication of both chronic and acute liver diseases. The term hepatic encephalopathy stems from the belief that hepatic insufficiency is its fundamental etiopathogenic factor. However, most clinical cases show liver failure along with mesenteric venous portal hypertension. This portal hypertension would explain the abnormal mechanical forces suffered by the digestive tract in the early stages of the disorder. These forces could regulate some gut biochemical pathological pathways in a process known as mechanotransduction. Thus, portal hypertension would begin with the establishment of a mechanotransduced afferent or sensory inflammatory gut-brain pathway, resulting in functional and structural changes in the central nervous system. In this review, we will revisit the term "hepatic encephalopathy" in light of new results where portal hypertension occurs before liver failure and is accompanied by brain changes. Moreover, we will point out cellular links that can explain the microbiota, immune, gut, and brain axis disturbances found in this disorder.

A Cellularized Biphasic Implant Based on a Bioactive Silk Fibroin Promotes Integration and Tissue Organization during Osteochondral Defect Repair in a Porcine Model.

In cartilage tissue engineering, biphasic scaffolds (BSs) have been designed not only to influence the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone, promoting the implant's integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a BS based on the assembly of a cartilage phase constituted by fibroin biofunctionalyzed with a bovine cartilage matrix, cellularized with differentiated autologous pre-chondrocytes and well attached to a bone phase (decellularized bovine bone) to promote cartilage regeneration in a model of joint damage in pigs. BSs were assembled by fibroin crystallization with methanol, and the mechanical features and histological architectures were evaluated. The scaffolds were cellularized and matured for 12 days, then implanted into an osteochondral defect in a porcine model (n = 4). Three treatments were applied per knee: Group I, monophasic cellular scaffold (single chondral phase); group II (BS), cellularized only in the chondral phase; and in order to study the influence of the cellularization of the bone phase, Group III was cellularized in chondral phases and a bone phase, with autologous osteoblasts being included. After 8 weeks of surgery, the integration and regeneration tissues were analyzed via a histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular BSs reached a Young's modulus of 805.01 kPa, similar to native cartilage. In vitro biological studies revealed the chondroinductive ability of the BSs, evidenced by an increase in sulfated glycosaminoglycans and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, in Group I, the defects were not reconstructed. In Groups II and III, a good integration of the implant with the surrounding tissue was observed. Defects in group II were fulfilled via hyaline cartilage and normal bone. Group III defects showed fibrous repair tissue. In conclusion, our findings demonstrated the efficacy of a biphasic and bioactive scaffold based on silk fibroin and cellularized only in the chondral phase, which entwined chondroinductive features and a biomechanical capability with an appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Current State of Transcranial Magnetic Stimulation and its use in Psychiatry.

INTRODUCTION: Transcranial magnetic stimulation (TMS) is a noninvasive brain stimulation technique that could be used as a therapeutic intervention in order to treat psychiatric disorders. AIM: Reviewing the effectiveness of TMS in the modulation of cognitive functions and also detailing its potential applications in psychiatric treatments. DEVELOPMENT: TMS has been traditionally used for the treatment of a great variety of neurological or psychiatric conditions by modulating the activity in brain areas and networks. Therapeutic benefit has been found in depressive disorders, anxiety, schizophrenia, addiction, and neurodevelopmental disorders as well as in brain damage and neurodegenerative disorders. Moreover, TMS is a technique which offers great tolerance and can be used as complement with other therapies. However, it is not easy to define an optimal treatment for every pathology: the parameters of stimulation are variable, and its effects at the cellular level of the nervous system are not well-known. CONCLUSION: While it is true that TMS provides many therapeutic benefits, it requires further investigation. It is necessary to detail the action mechanism of the stimulation and the long-term side effects, if any. This information would allow the design of specific treatment protocols for different psychiatric disorders.

Why are maternally separated females inflexible? Brain activity pattern of COx and c-Fos.

Subjects' early life events will affect them later in life. When these events are stressful, such as child abuse in humans or repeated maternal separation in rodents, subjects can show some behavioral and brain alterations. This study used young adult female Wistar rats that were maternally raised (AFR), maternally separated from post-natal day (PND) 1 to PND10 (MS10), or maternally separated from PND1 to PND21 (MS21), in order to assess the effects of maternal separation (MS) on spatial learning and memory, as well as cognitive flexibility, using the Morris Water Maze (MWM). We performed quantitative cytochrome oxidase (COx) histochemistry on selected brain areas in order to identify whether maternal separation affects brain energy metabolism. We also performed c-Fos immunohistochemistry on the medial prefrontal cortex (mPFC), thalamus, and hippocampus to explore whether this immediate early gene activity was altered in stressed subjects. We obtained a similar spatial learning pattern in maternally raised and maternally separated subjects on the reference memory task, but only the controls were flexible enough to solve the reversal learning successfully. Separated groups showed less c-Fos activity in the mPFC and less complex neural networks on COx.

Lobomycosis-like disease in common bottlenose dolphins Tursiops truncatus from Belize and Mexico: bridging the gap between the Americas.

Lobomycosis and lobomycosis-like diseases (LLD) (also: paracoccidioidomycosis) are chronic cutaneous infections that affect Delphinidae in tropical and subtropical regions worldwide. In the Americas, these diseases have been relatively well-described, but gaps still exist in our understanding of their distribution across the continent. Here we report on LLD affecting inshore bottlenose dolphins Tursiops truncatus from the Caribbean waters of Belize and from the eastern tropical Pacific Ocean off the southwestern coast of Mexico. Photo-identification and catalog data gathered between 1992 and 2017 for 371 and 41 individuals, respectively from Belize and Mexico, were examined for the presence of LLD. In Belize, 5 free-ranging and 1 stranded dolphin were found positive in at least 3 communities with the highest prevalence in the south. In Guerrero, Mexico, 4 inshore bottlenose dolphins sighted in 2014-2017 were affected by LLD. These data highlight the need for histological and molecular studies to confirm the etiological agent. Additionally, we document a single case of LLD in an adult Atlantic spotted dolphin Stenella frontalis in southern Belize, the first report in this species. The role of environmental and anthropogenic factors in the occurrence, severity, and epidemiology of LLD in South and Central America requires further investigation.

Dynamics of Goldfish Subregional Hippocampal Pallium Activity throughout Spatial Memory Formation.

The teleost fish hippocampal pallium, like the hippocampus of tetrapods, is essential for relational map-like spatial memories. In mammals, these relational memories involve the dynamic interactions among different hippocampal subregions and between the hippocampus-neocortex network, which performs specialized operations such as memory encoding and retrieval. However, how the teleost hippocampal homologue operates to achieve comparably sophisticated spatial cognition capabilities is largely unknown. In the present study, the progressive changes in the metabolic activity of the pallial regions that have been proposed as possible homologues of the mammalian hippocampus were monitored in goldfish. Quantitative cytochrome oxidase histochemistry was used to measure the level of activation along the rostrocaudal axis of the ventral (Dlv) and dorsal parts of the dorsolateral division (Dld) and in the dorsoposterior division (Dp) of the goldfish telencephalic pallium throughout the time course of the learning process of a spatial memory task. The results revealed a significant increase in spatial memory-related metabolic activity in the Dlv, but not in the Dld, suggesting that the Dlv, but not the Dld, is comparable to the amniote hippocampus. Regarding the Dlv, the level of activation of the precommissural Dlv significantly increased at training onset but progressively declined to finally return to the basal pretraining level when the animals mastered the spatial task. In contrast, the commissural Dlv activation persisted even when the acquisition phase was completed and the animal's performance reached an asymptotic level. These results suggest that, like the dentate gyrus of mammals, the goldfish precommissural Dlv seems to respond nonlinearly to increments of change in sensory input, performing pattern separation under highly dissimilar input patterns. In addition, like the CA3 of mammals, the commissural Dlv likely operates in a continuum between two modes, a pattern separation or storage operation mode at early acquisition when the change in the sensory input is high, probably driven by the precommissural Dlv output, and a pattern completion or recall operation mode when the animals have mastered the task and the change in sensory input is small. Finally, an unexpected result of the present study is the persistent activation of the area Dp throughout the complete spatial task training period, which suggests that the Dp could be an important component of the pallial network involved in spatial memory in goldfish, and supports the hypothesis proposing that the Dp is a specialized part of the hippocampal pallium network.