Species Composition and Feeding Behaviors of Vector Mosquitoes of Avian Infectious Diseases at a Wild Bird Rehabilitation Facility in Japan.

Although wild bird rehabilitation facilities are important for the conservation of wild species, individuals may be kept within the facilities for long periods, consequently posing a risk for the bird to be infected with pathogens to which they are not naturally exposed. In turn, novel pathogens may be introduced through rescued migratory species. Avian malaria and West Nile fever are important avian diseases transmitted by mosquitoes. To understand the transmission dynamics of such diseases at rehabilitation facilities, the ecology of vector mosquitoes, including species composition, seasonality, and feeding behaviors, were explored. Mosquitoes were collected at a wild bird rehabilitation facility and wildlife sanctuary in Japan from 2019 to 2020 using mouth aspirators, sweep nets, and light traps. A total of 2,819 mosquitoes of 6 species were captured, all of which are potential vectors of avian diseases. Culex pipiens pallens and Cx. pipiens form molestus were the dominant species (82.9% of all collected mosquitoes). Density and seasonality differed between sampling locations, presumably because of differences in mosquito behaviors including feeding preferences and responses to climatic factors. Blood-fed Culex mosquitoes fed solely on birds, and many mosquito species are thought to have fed on birds within the facility. Particularly, Cx. pipiens group probably fed on both rescued and free-living birds. The rehabilitation facility may be an important site for the introduction and spread of pathogens because 1) numerous mosquitoes inhabit the hospital and its surroundings; 2) blood-fed mosquitoes are caught within the hospital; 3) there is direct contact between birds and mosquitoes; 4) both birds within the hospital and wild birds are fed upon. Furthermore, blood-fed Cx. pipiens form molestus were observed in the winter, suggesting that pathogens might be transmitted even during the winter when other mosquito species are inactive.

Encoding of self-initiated actions in axon terminals of the mesocortical pathway.

Significance: The initiation of goal-directed actions is a complex process involving the medial prefrontal cortex and dopaminergic inputs through the mesocortical pathway. However, it is unclear what information the mesocortical pathway conveys and how it impacts action initiation. In this study, we unveiled the indispensable role of mesocortical axon terminals in encoding the execution of movements in self-initiated actions. Aim: To investigate the role of mesocortical axon terminals in encoding the execution of movements in self-initiated actions. Approach: We designed a lever-press task in which mice internally determine the timing of the press, receiving a larger reward for longer waiting periods. Results: Our study revealed that self-initiated actions depend on dopaminergic signaling mediated by D2 receptors, whereas sensory-triggered lever-press actions do not involve D2 signaling. Microprism-mediated two-photon calcium imaging further demonstrated ramping activity in mesocortical axon terminals approximately 0.5 s before the self-initiated lever press. Remarkably, the ramping patterns remained consistent whether the mice responded to cues immediately for a smaller reward or held their response for a larger reward. Conclusions: We conclude that mesocortical dopamine axon terminals encode the timing of self-initiated actions, shedding light on a crucial aspect of the intricate neural mechanisms governing goal-directed behavior.

Functional diversity of dopamine axons in prefrontal cortex during classical conditioning.

Midbrain dopamine neurons impact neural processing in the prefrontal cortex (PFC) through mesocortical projections. However, the signals conveyed by dopamine projections to the PFC remain unclear, particularly at the single-axon level. Here, we investigated dopaminergic axonal activity in the medial PFC (mPFC) during reward and aversive processing. By optimizing microprism-mediated two-photon calcium imaging of dopamine axon terminals, we found diverse activity in dopamine axons responsive to both reward and aversive stimuli. Some axons exhibited a preference for reward, while others favored aversive stimuli, and there was a strong bias for the latter at the population level. Long-term longitudinal imaging revealed that the preference was maintained in reward- and aversive-preferring axons throughout classical conditioning in which rewarding and aversive stimuli were paired with preceding auditory cues. However, as mice learned to discriminate reward or aversive cues, a cue activity preference gradually developed only in aversive-preferring axons. We inferred the trial-by-trial cue discrimination based on machine learning using anticipatory licking or facial expressions, and found that successful discrimination was accompanied by sharper selectivity for the aversive cue in aversive-preferring axons. Our findings indicate that a group of mesocortical dopamine axons encodes aversive-related signals, which are modulated by both classical conditioning across days and trial-by-trial discrimination within a day.

Functional Diversity of Dopamine Axons in Prefrontal Cortex During Classical Conditioning.

Midbrain dopamine neurons impact neural processing in the prefrontal cortex (PFC) through mesocortical projections. However, the signals conveyed by dopamine projections to the PFC remain unclear, particularly at the single-axon level. Here, we investigated dopaminergic axonal activity in the medial PFC (mPFC) during reward and aversive processing. By optimizing microprism-mediated two-photon calcium imaging of dopamine axon terminals, we found diverse activity in dopamine axons responsive to both reward and aversive stimuli. Some axons exhibited a preference for reward, while others favored aversive stimuli, and there was a strong bias for the latter at the population level. Long-term longitudinal imaging revealed that the preference was maintained in reward- and aversive-preferring axons throughout classical conditioning in which rewarding and aversive stimuli were paired with preceding auditory cues. However, as mice learned to discriminate reward or aversive cues, a cue activity preference gradually developed only in aversive-preferring axons. We inferred the trial-by-trial cue discrimination based on machine learning using anticipatory licking or facial expressions, and found that successful discrimination was accompanied by sharper selectivity for the aversive cue in aversive-preferring axons. Our findings indicate that a group of mesocortical dopamine axons encodes aversive-related signals, which are modulated by both classical conditioning across days and trial-by-trial discrimination within a day.

Rectus capitis lateralis muscle revisited: a histological study using human fetuses.

BACKGROUND: Rectus capitis lateralis muscle (RCLM) is located at the border between the ventral and dorsal muscle groups, but the nerve topographical anatomy around the muscle is obscure. MATERIALS AND METHODS: We observed the RCLM in histological sections of 12 midterm and 10 near-term fetal heads (9-18 and 26-40 weeks of gestational age). RESULTS: At midterm, the RCLM wrapped around the inferiorly protruding inferolateral corner of the cartilaginous occipital bone. The muscle was adjacent to, or even continued to, the intertransversarius muscle between the atlas and axis. At near-term, the jugular process of the occipital bone, that is, the RCLM upper insertion, was either cartilaginous or bony, depending on age. The process formed a collar supporting the internal jugular vein from the inferior side. Moreover, the muscle is tightly attached to or inserted into the venous wall itself. The cartilaginous jugular process was adjacent to Reichert's cartilage, and the uppermost muscle fibers passed through a narrow space between these cartilages. The RCLM appeared to accelerate the jugular process elongation, resulting in complete union of the occipital and temporal bones. The ventral ramus of the first cervical nerve passed between the RCLM and rectus capitis anterior muscle to reach the longus capitis muscle. No nerve passed between the RCLM and the obliquus capitis superior muscle (a muscle at the suboccipital triangle). CONCLUSION: The dorsoventral position of the RCLM seemed to correspond to the scalenus posterior muscle in a laminar arrangement of the cervical axial musculature.

A Canonical Scheme of Bottom-Up and Top-Down Information Flows in the Frontoparietal Network.

Goal-directed behavior often involves temporal separation and flexible context-dependent association between sensory input and motor output. The control of goal-directed behavior is proposed to lie in the frontoparietal network, but the computational architecture of this network remains elusive. Based on recent rodent studies that measured and manipulated projection neurons in the frontoparietal network together with findings from earlier primate studies, we propose a canonical scheme of information flows in this network. The parietofrontal pathway transmits the spatial information of a sensory stimulus or internal motor bias to drive motor programs in the frontal areas. This pathway might consist of multiple parallel connections, each controlling distinct motor effectors. The frontoparietal pathway sends the spatial information of cognitively processed motor plans through multiple parallel connections. Each of these connections could support distinct spatial functions that use the motor target information, including attention allocation, multi-body part coordination, and forward estimation of movement state (i.e., forward models). The parallel pathways in the frontoparietal network enable dynamic interactions between regions that are tuned for specific goal-directed behaviors. This scheme offers a promising framework within which the computational architecture of the frontoparietal network and the underlying circuit mechanisms can be delineated in a systematic way, providing a holistic understanding of information processing in this network. Clarifying this network may also improve the diagnosis and treatment of behavioral deficits associated with dysfunctional frontoparietal connectivity in various neurological disorders including Alzheimer's disease.

Fetal development and growth of the human erector spinae with special reference to attachments on the surface aponeurosis.

PURPOSE: The longissimus (LO) and iliocostalis (IC) of adults consist of myofibers extending from the superolateral to the inferomedial side of the back and, because of the same course, they are fused in the thoracolumbar region. The LO also has a medial attachment to the long myofibers of the transversospinalis (TS) showing a course from the superomedial to the inferolateral side. However, there is apparently no information regarding when and how these similar longitudinal muscles differentiate from a cluster of dorsomedial myotome cells. METHODS: We examined sagittal and horizontal sections of the trunks of 39 human embryos and fetuses (18-330 mm crown-rump length). RESULTS: At 6-7 weeks gestational age (GA), the surface aponeurosis appeared prior to and independent of the thoracolumbar fascia. At 6-9 weeks GA, the LO myofibers had a postero-inferior course, from the transverse process to the initial aponeurosis, whereas the TS myofibers had a postero-superior course, from a lateral extension of the intertransverse ligament to the aponeurosis. However, the IC consisted of supracostal longitudinal myofibers and was distant from the LO until 12 weeks GA. Because of the lack of ligamentous attachments and ribs, myofibers of the TS, LO, and IC took a similar inferior course in the lumbar region. When the early TS was represented by the transverso-aponeurotic muscle, consequently, the LO corresponded to the aponeuro-transversal muscle and was independent from the IC. CONCLUSION: The classical model of TS and LO development does not recognize the essential role of the aponeurosis identified here.

Long-range connections enrich cortical computations.

The cerebral cortex can perform powerful computations, including those involved in higher cognitive functions. Cortical processing for such computations is executed by local circuits and is further enriched by long-range connectivity. This connectivity is activated under specific conditions and modulates local processing, providing flexibility in the computational performance of the cortex. For instance, long-range connectivity in the primary visual cortex exerts facilitatory impacts when the cortex is silent but suppressive impacts when the cortex is strongly sensory-stimulated. These dual impacts can be captured by a divisive gain control model. Recent methodological advances such as optogenetics, anatomical tracing, and two-photon microscopy have enabled neuroscientists to probe the circuit and synaptic bases of long-range connectivity in detail. Here, I review a series of evidence indicating essential roles of long-range connectivity in visual and hierarchical processing involving numerous cortical areas. I also describe an overview of the challenges encountered in investigating underlying synaptic mechanisms and highlight recent technical approaches that may overcome these difficulties and provide new insights into synaptic mechanisms for cortical processing involving long-range connectivity.

Host-parasite relationships between seabirds and the haemadipsid leech Chtonobdella palmyrae (Annelida: Clitellata) inhabiting oceanic islands in the Pacific Ocean.

The duognathous haemadipsid leeches of the genus Chtonobdella show a trans-oceanic distribution throughout the Indo-Pacific region. Although passive long-distance dispersal (LDD) of Chtonobdella leeches by birds has been suggested, little is known about the host-parasite relationships between avian hosts and Chtonobdella leeches. In the current study, we investigated Chtonobdella leech infestations of the eyes and other mucus membranes of migratory procellariiform seabirds, Pterodroma hypoleuca and Oceanodroma tristrami, captured at six locations in the Bonin Islands, Honshu and Okinawa Island, Japan. Analyses of the partial sequences of 18S rRNA, 28S rRNA, and mitochondrial cytochrome c oxidase subunit I (COI) and morphological examination of the specimens demonstrated that the Chtonobdella leeches belonged to Chtonobdella palmyrae, which is indigenous to Palmyra Atoll in the Northern Line Islands. A dominant COI sequence type was observed in samples from all six sites; therefore, C. palmyrae almost surely dispersed approximately 1000 km by infesting the eyes and mucus membranes of procellariiform seabirds. The host-parasite relationships between procellariiform seabirds and C. palmyrae provide explicit evidence of the LDD of duognathous haemadipsid leeches. The taxonomic status of Haemadipsa zeylanica ivosimae from the Volcano Islands is also briefly discussed.

The effects of migration on the immunity of Black-Headed Gulls (Chroicocephalus ridibundus: Laridae).

In order to elucidate the relationship between migration period and immunity related to susceptibility, we conducted research on Black-headed gulls (Chroicocephalus ridibundus). We captured 260 gulls and collected their peripheral blood. Their leukocyte (WBC) count, percentages of heterophils (Het) and lymphocytes (Lym), heterophil and lymphocyte ratio (H/L ratio), and CD4 and CD8α expression levels (CD4 and CD8α, respectively) were quantitatively analyzed over three migration periods (Autumn migration, Wintering, Spring migration). In Adult gulls, WBC counts and CD4 levels significantly increased. Moreover, the Het and H/L ratio decreased from the Autumn migration to Wintering. Conversely, only WBC counts and CD4 levels measurements significantly decreased from Wintering to Spring migration (P<0.05). The tested parameters of the Tokyo-bay population show a greater significant difference than the measurements of immunity of the Mikawa-bay population. This study suggests that the migratory period has a negative effect on an aspect of the immune system. Including the period-difference in the immune systems in the local population, it is necessary to investigate the relationship between the ecology of migratory birds and their immunity.

Relationship between blood test values and blood lead (Pb) levels in Black-headed gull (Chroicocephalus ridibundus: Laridae).

Few studies have evaluated immunosuppression due to lead accumulation below the overt toxicity threshold. If low levels of lead accumulation cause immunosuppression in birds, those birds could become more susceptible to pathogens. We aimed to determine if low levels of lead accumulation lead to immunosuppression in Black-headed gulls (Chroicocephalus ridibundus). Gulls were captured in Tokyo-bay and Mikawa-bay from January to April 2019. Their blood samples were analyzed for eight items. The data were analyzed to evaluate the correlation between lead concentrations and the variables from each bay. Lead was positively correlated with the percentage of heterophils and heterophil and lymphocyte ratio and negatively with lymphocytes. Thus, low lead accumulation levels may induce changes in percentage of the heterophils and lymphocyte.

Interhemispherically dynamic representation of an eye movement-related activity in mouse frontal cortex.

Cortical plasticity is fundamental to motor recovery following cortical perturbation. However, it is still unclear how this plasticity is induced at a functional circuit level. Here, we investigated motor recovery and underlying neural plasticity upon optogenetic suppression of a cortical area for eye movement. Using a visually-guided eye movement task in mice, we suppressed a portion of the secondary motor cortex (MOs) that encodes contraversive eye movement. Optogenetic unilateral suppression severely impaired contraversive movement on the first day. However, on subsequent days the suppression became inefficient and capability for the movement was restored. Longitudinal two-photon calcium imaging revealed that the regained capability was accompanied by an increased number of neurons encoding for ipsiversive movement in the unsuppressed contralateral MOs. Additional suppression of the contralateral MOs impaired the recovered movement again, indicating a compensatory mechanism. Our findings demonstrate that repeated optogenetic suppression leads to functional recovery mediated by the contralateral hemisphere.

Streamlined sensory motor communication through cortical reciprocal connectivity in a visually guided eye movement task.

Cortical computation is distributed across multiple areas of the cortex by networks of reciprocal connectivity. However, how such connectivity contributes to the communication between the connected areas is not clear. In this study, we examine the communication between sensory and motor cortices. We develop an eye movement task in mice and combine it with optogenetic suppression and two-photon calcium imaging techniques. We identify a small region in the secondary motor cortex (MOs) that controls eye movements and reciprocally connects with a rostrolateral part of the higher visual areas (VRL/A/AL). These two regions encode both motor signals and visual information; however, the information flow between the regions depends on the direction of the connectivity: motor information is conveyed preferentially from the MOs to the VRL/A/AL, and sensory information is transferred primarily in the opposite direction. We propose that reciprocal connectivity streamlines information flow, enhancing the computational capacity of a distributed network.

Analysis of body mass changes in the Black-Headed Gull (Larus ridibundus) during the winter.

During the autumn migration of many waterfowls, body mass is lowest upon arrival at the wintering area and gradually increases during the winter. Consequently, body mass is highest before the spring migration. We studied the pattern of body mass changes in the Black-Headed Gull (Larus ridibundus) from December 2010 to December 2016 in the Shinhama area of Chiba, Japan. Based on 327 captured animals, body mass increased during the wintering period, but tended to decrease before migration. In 2014-2016, a muscle mass reduction in females was observed, explaining the change in body mass. However, the observed weight loss may be caused by many factors, which may be related to the migratory ecology of the regional population.

Selective Suppression of Local Circuits during Movement Preparation in the Mouse Motor Cortex.

Prepared movements are more efficient than those that are not prepared for. Although changes in cortical activity have been observed prior to a forthcoming action, the circuits involved in motor preparation remain unclear. Here, we use in vivo two-photon calcium imaging to uncover changes in the motor cortex during variable waiting periods prior to a forepaw reaching task in mice. Consistent with previous reports, we observed a subset of neurons with increased activity during the waiting period; however, these neurons did not account for the degree of preparation as defined by reaction time (RT). Instead, the suppression of activity of distinct neurons in the same cortical area better accounts for RT. This suppression of neural activity resulted in a distinct and reproducible pattern when mice were well prepared. Thus, the selective suppression of network activity in the motor cortex may be a key feature of prepared movements.

Influence of Si Addition on Quenching Sensitivity and Formation of Nano-Precipitate in Al-Mg-Si Alloys.

The age-hardening is enhanced with the high cooling rate since more vacancies are formed during quenching, whereas the stable beta phase is formed with the slow cooling rate just after solid solution treatment resulting in lower increase of hardness during aging. Meanwhile, the nanoclusters are formed during natural aging in Al-Mg-Si alloys. The formation of nanoclusters is enhanced with increasing the Si amount. High quench sensitivity based on mechanical property changes was confirmed with increasing the Si amount. Moreover, the nano-size beta" phase, main hardening phase, is more formed by the Si addition resulting in enhancement of the age-hardening. The quench sensitivity and the formation behavior of precipitates are discussed based on the age-hardening phenomena.

The Orientation and Variation of the Acromioclavicular Ligament: An Anatomic Study.

BACKGROUND: Several biomechanical studies have shown that the acromioclavicular (AC) ligament prevents posterior translation of the clavicle in the horizontal plane. In anatomy textbooks, however, the AC ligament is illustrated as running straight across the AC joint surface. HYPOTHESIS: The AC ligament does not run straight across the joint surface, and the configuration of the AC ligament may vary. STUDY DESIGN: Descriptive laboratory study. METHODS: We used 16 pairs of shoulder girdles in this study. After identifying the AC ligament, we macroscopically investigated the orientation and attachment of the ligament and measured the angle between the ligament and the line perpendicular to the AC joint surface by using a digital goniometer. In addition, the AC joint inclination angle was measured, and the Spearman rank correlation coefficient between the joint inclination and the ligament angle was calculated. Finally, we sought to classify the AC ligament based on its configuration. Of the 16 pairs of specimens, 3 pairs of shoulders were histologically examined. RESULTS: The AC ligament was divided into 2 parts: a bundle at the superoposterior (SP) part and a bundle at the anteroinferior (AI) part of the joint. The well-developed SP bundle was consistent and ran obliquely at an average ± SD 30° ± 6° in relation to the AC joint surface, from the anterior part of the acromion to the posterior part of the distal clavicle. The joint inclination was 70° ± 12°, and a negative moderate correlation was found between the joint inclination and the ligament angle (P = .02, r = -0.46). In comparison, the AI bundle was thin and narrow, and it could be categorized into 3 types according to its various configurations. CONCLUSION: The AC ligament could be separated into the SP bundle and the AI bundle. The SP bundle ran posteriorly toward the distal clavicle from the acromion at an average angle of 30° to the joint surface. CLINICAL RELEVANCE: Anatomic reconstruction, based on the current findings in combination with findings regarding the coracoclavicular ligament, could facilitate improved outcome in the treatment of AC joint disruption.

An excitatory basis for divisive normalization in visual cortex.

Neurons in visual cortex are connected not only locally, but also through networks of distal connectivity. These distal networks recruit both excitatory and inhibitory synapses and result in divisive normalization. Normalization is traditionally thought to result from increases in synaptic inhibition. By combining optogenetic stimulation and intracellular recordings in mouse visual cortex, we found that, on the contrary, normalization is a result of a decrease in synaptic excitation.

Imaging the awake visual cortex with a genetically encoded voltage indicator.

Genetically encoded voltage indicators (GEVIs) promise to reveal the membrane potential of genetically targeted neuronal populations through noninvasive, chronic imaging of large portions of cortical space. Here we test a promising GEVI in mouse cortex during wakefulness, a challenging condition due to large hemodynamic activity, and we introduce a straightforward projection method to separate a signal dominated by membrane voltage from a signal dominated by hemodynamic activity. We expressed VSFP-Butterfly 1.2 plasmid in layer 2/3 pyramidal cells of visual cortex through electroporation in utero. We then used wide-field imaging with two cameras to measure both fluorophores of the indicator in response to visual stimuli. By taking weighted sums and differences of the two measurements, we obtained clear separation of hemodynamic and voltage signals. The hemodynamic signal showed strong heartbeat oscillations, superimposed on slow dynamics similar to blood oxygen level-dependent (BOLD) or "intrinsic" signals. The voltage signal had fast dynamics similar to neural responses measured electrically, and showed an orderly retinotopic mapping. We compared this voltage signal with calcium signals imaged in transgenic mice that express a calcium indicator (GCaMP3) throughout cortex. The voltage signal from VSFP had similar signal-to-noise ratios as the calcium signal, it was more immune to vascular artifacts, and it integrated over larger regions of visual space, which was consistent with its reporting mostly subthreshold activity rather than the spiking activity revealed by calcium signals. These results demonstrate that GEVIs provide a powerful tool to study the dynamics of neural populations at mesoscopic spatial scales in the awake cortex.

Effect of corn steep liquor on lettuce root rot (Fusarium oxysporum f.sp. lactucae) in hydroponic cultures.

BACKGROUND: Recent reports indicate that organic fertilisers have a suppressive effect on the pathogens of plants grown under hydroponic systems. Furthermore, microorganisms exhibiting antagonistic activity to diseases have been observed in organic hydroponic systems. This study evaluated the effect of corn steep liquor (CSL) on controlling lettuce root rot disease [Fusarium oxysporum f.sp. lactucae (FOL)] in a hydroponic system. The effect of CSL and Otsuka A (a chemical fertiliser) on the inhibition of FOL in terms of mycelial growth inhibition was tested in vivo. RESULTS: Addition of CSL suppressed FOL infection rates. CSL inhibited FOL infection by 26.3-42.5% from 2 days after starting incubation. In comparison, Otsuka A inhibited FOL growth by 5.5-19.4%. In addition, four of 10 bacteria isolated from the nutrient media containing CSL exhibited inhibition zones preventing FOL mycelial growth. CONCLUSIONS: We found that CSL suppressed FOL in lettuce via its antifungal and biostimulatory effects. We suggest that activation of beneficial microorganisms present in CSL may be used to decrease lettuce root rot disease and contribute to lettuce root growth.