Weighted gene co-expression network analysis reveals immune evasion related genes in Echinococcus granulosus sensu stricto.

Cystic echinococcosis (CE) is a zoonotic disease caused by the tapeworm Echinococcus granulosus sensu lato (s.l). In the intermediate host, this disease is characterized by the growth of cysts in viscera such as liver and lungs, inside of which the parasite develops to the next infective stage known as protoscoleces. There are records that the infected viscera affect the development and morphology of E. granulosus s.l. protoscolex in hosts such as buffalo or humans. However, the molecular mechanisms that drive these differences remains unknown. Weighted gene co-expression network analysis (WGCNA) using a set of RNAseq data obtained from E. granulosus sensu stricto (s.s.) protoscoleces found in liver and lung cysts reveals 34 modules in protoscoleces of liver origin, of which 12 have differential co-expression from protoscoleces of lung origin. Three of these twelve modules contain hub genes related to immune evasion: tegument antigen, tegumental protein, ubiquitin hydrolase isozyme L3, COP9 signalosome complex subunit 3, tetraspanin CD9 antigen, and the methyl-CpG-binding protein Mbd2. Also, two of the twelve modules contain only hypothetical proteins with unknown orthology, which means that there are a group of unknown function proteins co-expressed inside the protoscolex of liver CE cyst origin. This is the first evidence of gene expression differences in protoscoleces from CE cysts found in different viscera, with co-expression networks that are exclusive to protoscoleces from liver CE cyst samples. This should be considered in the control strategies of CE, as intermediate hosts can harbor CE cysts in liver, lungs, or both organs simultaneously.

Molecular surveillance of potential SARS-CoV-2 reservoir hosts in wildlife rehabilitation centers.

BACKGROUND: The COVID-19 pandemic, caused by SARS-CoV-2 infection, has become the most devastating zoonotic event in recent times, with negative impacts on both human and animal welfare as well as on the global economy. Although SARS-CoV-2 is considered a human virus, it likely emerged from animals, and it can infect both domestic and wild animals. This constitutes a risk for human and animal health including wildlife with evidence of SARS-CoV-2 horizontal transmission back and forth between humans and wild animals. AIM: Molecular surveillance in different wildlife rehabilitation centers and wildlife associated institutions in Chile, which are critical points of animal-human interaction and wildlife conservation, especially since the aim of wildlife rehabilitation centers is to reintroduce animals to their original habitat. MATERIALS AND METHODS: The survey was conducted in six WRCs and three wildlife associated institutions. A total of 185 samples were obtained from 83 individuals belonging to 15 different species, including vulnerable and endangered species. Each specimen was sampled with two different swabs: one oropharyngeal or nasopharyngeal according to the nostril diameter, and/or a second rectal sample. RNA was extracted from the samples and two different molecular assays were performed: first, a conventional RT-PCR with pan-coronavirus primers and a second SARS-CoV-2 qPCR targeting the N and S genes. RESULTS: All 185 samples were negative for SARS-CoV-2. CLINICAL RELEVANCE: This study constitutes the first report on the surveillance of SARS-CoV-2 from wildlife treated in rehabilitation centers in Chile, and supports the biosafety procedures adopted in those centers.

Análisis biomecánico comparativo de recepción a media altura entre jugadores profesionales y amateurs de fútbol / Análise biomecânica comparativa da captura em meia altura entre jogadores profissionais e amadores de futebol / Comparative biomechanical analysis of mid-height reception between professional and amateur soccer players

RESUMEN El fútbol es un deporte de varias acciones motrices, donde la recepción del balón se clasifica como una acción de control a diferentes alturas, velocidades y ángulos. El jugador debe tener la capacidad de receptar el balón para poder generar una acción diferente: un pase, tiro o driblar; su optimización implica mayores probabilidades de rendimiento técnico-táctico. En tal sentido, se planteó como objetivo de la investigación analizar biomecánicamente la técnica de la recepción a media altura en futbolistas profesionales y amateurs a partir de video análisis como método de medición. La investigación fue descriptiva-explicativa de orden correlacional, se analizan cuatro variables del movimiento motriz (D: distancia Recorrida; AM: aceleración máxima; VM: velocidad máxima y AF: amplitud de la articulación femorotibial) a través de video-análisis con Kinovea. Se estudiaron a cinco jugadores profesionales (grupo 1) y 15 de nivel amateur (grupo 2). Los futbolistas de nivel amateur presentaron una mayor media en todas las variables estudiadas, "D" (grupo 1: 72.17cm; grupo 2: 101.27cm; p=0.001), "AM" (grupo 1: 61.84m/s2; grupo 2: 101.73m/s2; p=0.000), "VM" (grupo 1: 8.18m/s; grupo 2: 12.55m/s; p=0.000) y "AF" (grupo 1: 62.6°; grupo 2: 75.2°; p=0.002), lo cual se comporta significativamente diferentes en todos los casos. El análisis biomecánico realizado a futbolistas profesionales y amateurs, en la técnica de recepción del balón a media altura, concluye que en los jugadores amateur requieren un mayor perfeccionamiento técnico, independientemente de su experiencia en competencias.

RESUMO O futebol é um esporte de várias ações motoras, onde a recepção da bola é classificada como uma ação de controle em diferentes alturas, velocidades e ângulos. O jogador deve ter a capacidade de receber a bola para poder gerar uma ação diferente: um passe, um chute ou um drible; sua otimização implica em maiores probabilidades de desempenho técnico-tático. Neste sentido, o objetivo da pesquisa era analisar biomecanicamente a técnica de recepção em meia altura em jogadores profissionais e amadores de futebol utilizando a análise de vídeo como método de medição. A pesquisa foi descritiva-explicativa de ordem correlacional, quatro variáveis de movimento motor (D: distância coberta; MA: aceleração máxima; MV: velocidade máxima e FA: amplitude da articulação femorotibial) foram analisadas através de vídeo-análise com Kinovea. Foram estudados cinco jogadores profissionais (grupo 1) e 15 jogadores amadores (grupo 2). Os jogadores de futebol de nível amador apresentaram uma média maior em todas as variáveis estudadas, "D" (grupo 1: 72,17cm; grupo 2: 101,27cm; p=0,001), "AM" (grupo 1: 61,84m/s2; grupo 2: 101. 73m/s2; p=0,000), "VM" (grupo 1: 8,18m/s2; grupo 2: 12,55m/s; p=0,000) e "AF" (grupo 1: 62,6°; grupo 2: 75,2°; p=0,002), que se comportam de forma significativamente diferente em todos os casos. A análise biomecânica realizada sobre jogadores profissionais e amadores de futebol, na técnica de recepção de bola em altura média, conclui que os jogadores amadores requerem um maior aperfeiçoamento técnico, independentemente de sua experiência em competições.

ABSTRACT Soccer is a sport of various motor actions, where the reception of the ball is classified as a control action at different heights, speeds and angles. The player must have the ability to receive the ball in order to generate a different action: a pass, shot or dribble; its optimization implies greater probabilities of technical-tactical performance. In this sense, the objective of the research was to biomechanically analyze the mid-height reception technique in professional and amateur soccer players using video analysis as a measurement method. The research was descriptive-explanatory of correlational order, four variables of the motor movement (D: distance covered; AM: maximum acceleration; VM: maximum speed and AF: amplitude of the femorotibial joint) are analyzed through video-analysis with Kinovea . Five professional players (group 1) and 15 amateur players (group 2) were studied. The amateur soccer players presented a higher mean in all the variables studied, "D" (group 1: 72.17cm; group 2: 101.27cm; p=0.001), "AM" (group 1: 61.84m/s 2 ; group 2: 101.73m/s 2 ; p=0.000), "VM" (group 1: 8.18m/s; group 2: 12.55m/s; p=0.000) and "AF" (group 1: 62.6°; group 2 : 75.2°; p=0.002), which behaves significantly different in all cases. The biomechanical analysis carried out on professional and amateur soccer players, in the technique of receiving the ball at mid-height, concludes that amateur players require greater technical improvement, regardless of their experience in competitions.

Transcriptome analysis of Echinococcus granulosus sensu stricto protoscoleces reveals differences in immune modulation gene expression between cysts found in cattle and sheep.

Cystic Echinococcosis (CE), a zoonotic parasitic disease, is caused by the cestode Echinococcus granulosus sensu lato. CE inflicts severe damage in cattle, sheep, and human hosts worldwide. Fertile CE cysts are characterized by the presence of viable protoscoleces. These parasite forms are studied with minimal contamination with host molecules. Hosts, cattle and sheep, show differences in their CE cyst fertility. The effect of the host in protoscolex transcriptome is not known. We genotyped and performed transcriptomic analysis on sheep protoscoleces obtained from liver and lung CE cysts. The transcriptomic data of Echinococcus granulosus sensu stricto protoscoleces from 6 lung CE cysts and 6 liver CE cysts were Collected. For host comparison analysis, 4 raw data files belonging to Echinococcus granulosus sensu stricto protoscoleces from cattle liver CE cysts were obtained from the NCBI SRA database. Principal component and differential expression analysis did not reveal any statistical differences between protoscoleces obtained from liver or lung cysts, either within the same sheep or different sheep hosts. Conversely, there are significant differences between cattle and sheep protoscolex samples. We found differential expression of immune-related genes. In cattle, 7 genes were upregulated in protoscoleces from liver cysts. In sheep, 3 genes were upregulated in protoscoleces from liver and lung CE cysts. Noteworthy, are the differential expression of antigen B, tegument antigen, and arginase-2 in samples obtained from sheep CE cysts, and basigin in samples from cattle CE cysts. These findings suggest that the host species is an important factor involved in the differential expression of immune related genes, which in turn is possibly related to the fertility of Echinococcus granulosus sensu stricto cysts.

[Bilateral adrenal hemorrhage after severe COVID-19 infection. Report of one case]. / Hemorragia adrenal bilateral en paciente recuperado de neumonía COVID-19 Grave. Caso Radiológico.

We report a 47-year-old male with a severe disease caused by COVID-19, who required mechanical ventilation for 18 ays. During the hospital stay he received dexamethasone and anticoagulation with heparin. After discharge a new chest CT scan showed homogeneous hypo vascular enlargement of both adrenal glands, not present in the initial imaging studies. It was interpreted as a subacute bilateral adrenal hemorrhage. The patient remained stable and without any complaint.

A Unique Approach to Project-Based Learning (PjBL) in a Veterinary Anatomy Course.

INTRODUCTION: Project-based learning (PjBL) is a teaching methodology designed to engage students in solving real-world problems, acknowledging that students are active agents of their learning process. This methodology has historically been popular in architecture and industrial sciences; however, its use in teaching veterinary anatomy is scarcely published. METHODS: Using information and communication technologies, the PjBL methodology was implemented in a first-year veterinary anatomy course. The methodology included teamwork and the selection of a routine object in the veterinary clinic at the beginning of the academic semester. The project's goal was to analyze the object and associate it with both a domestic animal species and an anatomical region, along with making and presenting a video or a simulation model about the object. RESULTS: More than 80% of students prefer active learning classes compared to traditional classes. In addition, 66% and 86% of students indicate that PjBL allowed them to improve their understanding of theoretical content for the first and second years of post-implementation, respectively. Students' self-assessment indicates that more than 80% of the students (first and second year post-implementation) felt they were responsible for the execution of the project, able to conduct research, and able to develop autonomous learning skills. After 2 years of PjBL implementation, failure rates in the course decreased by 21%. DISCUSSION: In general, PjBL results show that veterinary students prefer active learning activities that allow them to learn in a team-based learning process as well as to develop soft skills such as self-learning, responsibility, and teamwork. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40670-021-01205-1.

Hemorragia adrenal bilateral en paciente recuperado de neumonía COVID-19 Grave: caso Radiológico / Bilateral adrenal hemorrhage after severe COVID-19 infection: report of one case

We report a 47-year-old male with a severe disease caused by COVID-19, who required mechanical ventilation for 18 ays. During the hospital stay he received dexamethasone and anticoagulation with heparin. After discharge a new chest CT scan showed homogeneous hypo vascular enlargement of both adrenal glands, not present in the initial imaging studies. It was interpreted as a subacute bilateral adrenal hemorrhage. The patient remained stable and without any complaint.

Response patterns in adventitial layer of Echinococcus granulosus sensu stricto cysts from naturally infected cattle and sheep.

Cystic echinococcosis is a zoonotic disease caused by the metacestode of Echinococcus granulosus sensu lato. The disease is characterized by the development of cystic structures inside viscera of the intermediate host, mainly liver and lungs. These cysts are formed by three layers: germinal, laminated, and adventitial layer, the latter being the local host immune response. Metacestodes that develop protoscoleces, the infective stage to the definitive host, are termed fertile, whereas cysts that do not produce protoscoleces are termed non-fertile. Sheep usually harbor fertile cysts while cattle usually harbor non-fertile cysts. Adventitial layers with fibrotic resolution are associated to fertile cysts, whereas a granulomatous reaction is associated with non-fertile cysts. The aim of this study was to analyze cellular distribution in the adventitial layer of fertile and non-fertile E. granulosus sensu stricto cysts found in liver and lungs of cattle and sheep. A total of 418 cysts were analyzed, 203 from cattle (8 fertile and 195 non-fertile) and 215 from sheep (64 fertile and 151 non-fertile). Fertile cysts from cattle showed mixed patterns of response, with fibrotic resolution and presence of granulomatous response in direct contact with the laminated layer, while sheep fertile cysts always displayed fibrotic resolution next to the laminated layer. Cattle non-fertile cysts display a granulomatous reaction in direct contact with the laminated layer, whereas sheep non-fertile cysts display a granulomatous reaction, but in direct contact with the fibrotic resolution. This shows that cattle and sheep cystic echinococcosis cysts have distinct local immune response patterns, which are associated to metacestode fertility.

Fasciola hepatica co-infection enhances Th1 immune response in the adventitial layer of non-fertile Echinococcus granulosus cysts.

Cystic echinococcosis is a widespread zoonosis caused by the larval stage of the tapeworm Echinococcus granulosus. In intermediary hosts, two types of echinococcal cysts can be found: fertile, which produce protoscoleces, the infective form of the parasite to dogs; and infertile, that do not present protoscoleces and therefore are not able to continue with the parasite life cycle. The adventitial layer, the local immune response against the cyst, plays an important role in cyst fertility. Grazing cattle can often feature Fasciola hepatica co-infection, a parasite known to modulate the host systemic immune response. In this work the cellular Th1/Th2 immune profiles were evaluated in the adventitial layer of fertile and non-fertile cysts with and without co-infection with Fasciola hepatica. Measuring with immunohistochemistry and qPCR in adventitial layer, we report that non-fertile cysts present higher levels of Th1 cytokines (IFN-γ (P < 0.0001) and TNF-α (P < 0.05)), and fertile cysts have higher levels of Th2 cytokines (IL-4 (P < 0.001)). Co-infection with Fasciola hepatica is associated with a decrease in the expression of IL-4 (P < 0.05) and an increase in the expression of IFN-γ (P < 0.0001) in the adventitial layer of fertile cysts. Non-fertile cysts were associated with higher levels of Th1 cytokines in the adventitial layer, with IFN-γ expression enhanced by F. hepatica co-infection (P < 0.0001), confirming that polyparasitism should be considered in the treatment and control of naturally infected cattle.

Cattle co-infection of Echinococcus granulosus and Fasciola hepatica results in a different systemic cytokine profile than single parasite infection.

E. granulosus is a cestode that causes Cystic Echinococcosis (CE), a zoonotic disease with worldwide presence. The immune response generated by the host against the metacestode induces a permissive Th2 response, as opposed to pro-inflammatory Th1 response. In this view, mixed Th2 and regulatory responses allow parasite survival. Overall, larval Echinococcus infections induce strong regulatory responses. Fasciola hepatica, another common helminth parasite, represents a major infection in cattle. Co-infection with different parasite species in the same host, polyparasitism, is a common occurrence involving E. granulosus and F. hepatica in cattle. 'While it is known that infection with F. hepatica also triggers a polarized Th2/Treg immune response, little is reported regarding effects on the systemic immune response of this example of polyparasitism. F. hepatica also triggers immune responses polarized to the Th2/ Treg spectrum. Serum samples from 107 animals were analyzed, and were divided according to their infection status and Echinococcal cysts fertility. Cytokines were measured utilizing a Milliplex Magnetic Bead Panel to detect IFN-γ, IL-1, IL-2, IL-4, IL-6, IL-10, IL-12 and IL-18. Cattle infected only with F. hepatica had the highest concentration of every cytokine analyzed, with both 4.24 and 3.34-fold increases in IL-10 and IL-4, respectively, compared to control animals, followed by E. granulosus and F. hepatica co-infected animals with two-fold increase in IL-10 and IL-4, compared to control animals, suggesting that E. granulosus co-infection dampens the cattle Th2/Treg immune response against F. hepatica. When considering Echinococcal cyst fertility and systemic cytokine concentrations, fertile cysts had higher IFN-γ, IL-6 and IL-18 concentrations, while infertile cysts had higher IL-10 concentrations. These results show that E. granulosus co-infection lowers Th1 and Th2 cytokine serological concentration when compared to F. hepatica infection alone. E. granulosus infections show no difference in IFN-γ, IL-1, IL-2, IL-6 and IL-18 levels compared with control animals, highlighting the immune evasion mechanisms of this cestode.

Fasciola hepatica coinfection modifies the morphological and immunological features of Echinococcus granulosus cysts in cattle.

Polyparasitism occurs when animals harbour multiple parasites concomitantly. It is a common occurrence but is generally understudied in wild and domestic animals. Fasciola hepatica and Echinococcus granulosus, which are helminths of ungulates, frequently coinfect cattle. The effects of this particular type of polyparasitism are not well documented. The metacestode of Echinococcus granulosus is surrounded by the adventitial layer, which constitutes the host immune response to the parasite. This layer in cattle is produced by a granulomatous reaction and is involved in echinococcal cyst (EC) fertility. Due to the systemic immune-modulating abilities of Fasciola hepatica, coinfection possibly generates a favourable environment for EC growth. A total of 203 Echinococcus granulosus sensu stricto cysts were found in 82 cattle, of which 42 ECs were found in 31 animals coinfected with Fasciola hepatica. The overall infection intensity was 3 cysts per animal. Coinfection with Fasciola hepatica decreased the mean infection intensity to 1.4 cysts per animal. Regarding EC size, coinfection resulted in smaller ECs (15.91 vs 22.09 mm), especially for infertile lung cysts. The adventitial layer of ECs in coinfected animals lacked lymphoid follicles and palisading macrophages, which are generally hallmarks of the granulomatous immune response. The ECs in coinfected animals had organized laminated layers, whereas those in animals without coinfection did not. Although coinfection was not statistically associated with EC fertility, we did not find fertile cysts in the livers of coinfected animals. We concluded that coinfection with Fasciola hepatica and Echinococcus granulosus has a detrimental effect on ECs, particularly infertile cysts.

Lymphocyte Populations in the Adventitial Layer of Hydatid Cysts in Cattle: Relationship With Cyst Fertility Status and Fasciola Hepatica Co-Infection.

Cystic echinococcosis is a worldwide zoonosis caused by the cestode Echinococcus granulosus. Two types of hydatid cysts occur in intermediate hosts: fertile cysts that generate protoscoleces from the germinal layer of the cyst, and infertile cysts that do not produce protoscoleces and are unable to continue the life cycle of the parasite. The adventitial layer, a host-derived fibrous capsule surrounding the hydatid cyst, is suggested to play an important role in local immune regulation during infection and in fertility of the cysts. Fasciola hepatica, another important parasite of cattle, induces a characteristic Th2-like immune response that could modulate the immune response against E. granulosus. Natural co-infection of both parasites is common in cattle, but no reports describe the local immune response against E. granulosus with F. hepatica infection in the same host. This study analyzed the number and distribution of T and B cells in the adventitial layer of liver and lung cysts and the relationship with cyst fertility and F. hepatica co-infection. T lymphocytes were the predominant cell type in the adventitial layer of infertile hydatid cysts and were more numerous in infertile hydatid cysts. B lymphocyte numbers were not associated with hydatid cyst fertility. Mast cells were infrequent in the adventitial layer. The number of T and B cells was not associated with F. hepatica co-infection. The present study contributes to the understanding of local immune responses in bovine cystic echinococcosis.

Multiple haplotypes of Echinococcus granulosus sensu stricto in single naturally infected intermediate hosts.

Cystic echinococcosis is a disease that affects both humans and animals, caused by cryptic species complex belonging to the platyhelminth Echinococcus granulosus sensu lato (s.l.). This disease is distributed worldwide, with E. granulosus sensu stricto (s.s.) being the most widespread of the species. High genetic variability has been demonstrated within E. granulosus s.s. studying single cyst per infected animal identifying a number of different haplotypes. However, few studies have addressed the genetic diversity of this parasite within a single intermediate host with multiple Echinococcus cysts. To date, it remains unknown if specific haplotypes of E. granulosus s.s. produce differences in biological features of the cyst. Here, we use the full length of the mitochondrial gene cox1 to determine E. granulosus s.s. haplotypes in samples from both cattle and sheep which harboured more than one cyst in different areas in Chile, where this parasite is endemic. We found 16 different haplotypes in 66 echinococcal cysts from 10 animals, and both cattle and sheep can harbour up to five different haplotypes of E. granulosus s.s. in the same animal. Regarding cyst fertility, five animals had both fertile and infertile Echinococcus cysts in both single and multiple haplotype infections. There was no association between haplotype and cyst fertility, size, or adventitial layer characteristics. Sampling and sequencing every Echinococcus cyst found in the intermediate host reveals a high molecular variability. We speculate that multiple haplotype infections could also suggest that intermediate hosts come from hyperendemic areas.

New insights of the local immune response against both fertile and infertile hydatid cysts.

BACKGROUND: Cystic echinococcosis is caused by the metacestode of the zoonotic flatworm Echinococcus granulosus. Within the viscera of the intermediate host, the metacestode grows as a unilocular cyst known as hydatid cyst. This cyst is comprised of two layers of parasite origin: germinal and laminated layers, and one of host origin: the adventitial layer, that encapsulates the parasite. This adventitial layer is composed of collagen fibers, epithelioid cells, eosinophils and lymphocytes. To establish itself inside the host, the germinal layer produces the laminated layer, and to continue its life cycle, generates protoscoleces. Some cysts are unable to produce protoscoleces, and are defined as infertile cysts. The molecular mechanisms involved in cyst fertility are not clear, however, the host immune response could play a crucial role. METHODOLOGY/PRINCIPAL FINDINGS: We collected hydatid cysts from both liver and lungs of slaughtered cattle, and histological sections of fertile, infertile and small hydatid cysts were stained with haematoxylin-eosin. A common feature observed in infertile cysts was the disorganization of the laminated layer by the infiltration of host immune cells. These infiltrating cells eventually destroy parts of laminated layer. Immunohistochemical analysis of both parasite and host antigens, identify these cells as cattle macrophages and are present inside the cysts associated to germinal layer. CONCLUSIONS/SIGNIFICANCE: This is the first report that indicates to cell from immune system present in adventitial layer of infertile bovine hydatid cysts could disrupt the laminated layer, infiltrating and probably causing the infertility of cyst.

Intra-articular treatment with corticosteroids increases apoptosis in human rotator cuff tears.

PURPOSE: The aim of this study is to evaluate in vivo the level of apoptosis in human rotator cuff tears and the relationship it might have with tendon degeneration. METHODS: Rotator cuff biopsies from 19 male and female patients, ages between 38 and 68 years, with and without previous corticosteroid infiltrations were collected via arthroscopy. Biopsies from seven patients with healthy rotator cuffs were used as a control group. An in situ terminal deoxynucleotidyl transferase dUTP nick end labeling assay was performed to detect the level of apoptosis, which was expressed as a percentage of apoptotic cells (PAC). RESULTS: PAC in patients with corticosteroid infiltrations was 76.97 ± 16.99 in all tendon rupture zones, in non-infiltrated patients was 35.89 ± 22.96, whereas in control patients was 14.48 ± 8.15. Likewise, the tendency of PAC reveals that apoptosis in control and non-infiltrated groups was different and dispersed in all tear zones; while in corticosteroid treated patients, the tendency was similar in all rupture sites. CONCLUSIONS: This investigation leads us to conclude that the administration of corticosteroid is associated with a higher amount of apoptosis at the insertion site of the rotator cuff (rupture edge).

Echinococcus granulosus hydatid cyst location is modified by Fasciola hepatica infection in cattle.

BACKGROUND: Natural parasite infection occurs in wild and domestics animals with more than one parasite species at the same time, generating an infection called polyparasitism. Cystic echinococcosis reports are usually based only on infection with Echinoccocus granulosus leaving aside other internal parasitoses that could modulate both the immune response and pathogenesis of the natural infection. Fasciola hepatica is another cosmopolitan parasite in ruminants with a similar distribution to E. granulosus in different parts of the world, but no information of the effect of co-infection with E. granulosus has been described. The aims of this report were to establish E. granulosus prevalence and explore the association of F. hepatica co-infection and natural E. granulosus infections in cattle. RESULTS: From 1725 animals, the prevalence of E. granulosus and F. hepatica was 21.16 and 51.3%, respectively. Considering both infections, older cattle (> 4 years) presented higher prevalence compared to younger animals. In E. granulosus-infected cattle, 5.21% had fertile cysts, 71.78% infertile cysts, and in 23.01% cysts were smaller than 1 cm in diameter. Considering cyst location, 39.72% had lungs cysts, 24.72% had liver cysts and 36.94% had cysts in both organs. Cyst location significantly differed between age groups: 44.68% of younger animals had cysts only in the lungs, while older animals presented hydatid cyst in the lungs and liver simultaneously (44.15%). With E. granulosus infection alone, 30.26% of cysts were found in the lungs, 31.79% in the liver and 37.95% in both organs. Regarding the co-infection of E. granulosus with F. hepatica, the proportion was significantly different (P < 0.05) with most animals having cysts only in the lungs (49.41%) and a lower level of liver infection (15.88%). Analyzing organ cyst distribution and F. hepatica absence/presence ratio within each cyst type, small cysts showed the highest difference in ratio. CONCLUSIONS: To the best of our knowledge, this is the first report indicating that F. hepatica co-infection in cattle could be affecting the instate of hydatid cysts in the liver, displacing toward lung localization, suggesting an antagonistic relationship.

First description of Echinococcus ortleppi and cystic echinococcosis infection status in Chile.

Cystic echinococcosis (CE), a parasitic disease caused by the cestode Echinococcus granulosus sensu lato (s.l.), is a worldwide zoonotic infection. Although endemic in Chile, information on the molecular characteristics of CE in livestock remains scarce. Therefore we aimed to describe the status of infection with E. granulosus s.l. in cattle from central Chile and also to contribute to the study of the molecular epidemiology of this parasite. According to our results, the prevalence of CE is 18.84% in cattle, similar to previous reports from Chile, suggesting that the prevalence in Santiago Metropolitan area has not changed in the last 30 years. Most of the cysts were found only in lungs (51%), followed by concurrent infection in liver and lungs (30%), and only liver (19%). Molecular characterization of the genetic diversity and population structure of E. granulosus s.l. from cattle in central Chile was performed using a section of the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene. E. granulosus sensu stricto (s.s.) (G1-G3 genotypes) was confirmed by RFLP-PCR to be the dominant species affecting cattle (284 samples/290 samples); we also report for the first time in Chile the presence of E. ortleppi (G5 genotype) (2 samples/61 samples). The Chilean E. granulosus s.s. parsimony network displayed 1 main haplotype. Additional studies using isolates from many locations across Chile and different intermediate hosts will provide more data on the molecular structure of E. granulosus s.s. within this region. Likewise, investigations of the importance of E. ortleppi in human infection in Chile deserve future attention.

Proteomics analysis of Echinococcus granulosus protoscolex stage.

Echinococcus granulosus protoscolex proteins were separated using two-dimensional electrophoresis and then identified using mass spectrometry; we identified 61 proteins, 28 which are newly described of which 4 could be involved in hydatid cyst fertility molecular mechanisms.

The TORC1/P70S6K and TORC1/4EBP1 signaling pathways have a stronger contribution on skeletal muscle growth than MAPK/ERK in an early vertebrate: Differential involvement of the IGF system and atrogenes.

Knowledge about the underlying mechanisms, particularly the signaling pathways that account for muscle growth in vivo in early vertebrates is still scarce. Fish (Paralichthys adspersus) were fasted for 3weeks to induce a catabolic period of strong muscle atrophy. Subsequently, fish were refed for 2weeks to induce compensatory muscle hypertrophy. During refeeding, the fish were treated daily with either rapamycin (TORC blocker), PD98059 (MEK blocker), or PBS (V; vehicle), or were untreated (C; control). Rapamycin and PD98059 differentially impaired muscle cellularity in vivo, growth performance, and the expression of growth-related genes, and the inhibition of TORC1 had a greater impact on fish muscle growth than the inhibition of MAPK. Blocking TORC1 inhibited the phosphorylation of P70S6K and 4EBP1, two downstream components activated by TORC1, thus affecting protein contents in muscle. Concomitantly, the gene expression in muscle of igf-1, 2 and igfbp-4, 5 was down-regulated while the expression of atrogin-1, murf-1, and igfbp-2, 3 was up-regulated. Muscle hypertrophy was abolished and muscle atrophy was promoted, which finally affected body weight. TORC2 complex was not affected by rapamycin. On the other hand, the PD98059 treatment triggered ERK inactivation, a downstream component activated by MEK. mRNA contents of igf-1 in muscle were down-regulated, and muscle hypertrophy was partially impaired. The present study provides the first direct data on the in vivo contribution of TORC1/P70S6K, TORC1/4EBP1, and MAPK/ERK signaling pathways in the skeletal muscle of an earlier vertebrate, and highlights the transcendental role of TORC1 in growth from the cellular to organism level.

Bovine IgG subclasses and fertility of Echinococcus granulosus hydatid cysts.

Hydatidosis is an important zoonotic disease of worldwide distribution, causing important health problems to humans and major economical losses in infected livestock. Echinococcus granulosus, the etiological agent of hydatid disease, induces a humoral immune response in the intermediate host (human and herbivorous) against hydatid cyst antigens. Specifically, IgGs are found in the laminar and germinal layers and inside the lumen of fertile and infertile hydatid cysts. In the germinal layer of infertile cysts IgGs are found in an order of magnitude greater than in the germinal layer of fertile cysts; a fraction of those IgGs are associated with high affinity to germinal layer proteins, suggesting their binding to specific parasite antigens. We have previously shown that those immunoglobulins, bound with high affinity to the germinal layer of hydatid cysts, induce apoptosis leading to cyst infertility. In the present work the presence of IgG1 and IgG2 subclasses in the germinal layer of both fertile and infertile hydatid cysts is reported. IgG1 is the most relevant immunoglobulin subclass present in the germinal layer of infertile cysts and bound with high affinity to that parasite structure. Contrarily, though the IgG2 subclass was also found in the germinal and adventitial layers, those immunoglobulins show low affinity to parasite antigens. We propose that the binding of an IgG1 subclass to parasite antigens present in the germinal layer is involved in the mechanism of cyst infertility.