Biochemical responses of Echinochloa polystachya inoculated with a Trichoderma consortium during the removal of a pyrethroid-based pesticide.

The biochemical response of plants exposed to pesticides and inoculated with microorganisms is of great importance to explore cleaning up strategies for contaminated sites with pyrethroid-based pesticides. We evaluated the effects of a Trichoderma consortium on the biochemical responses of Echinochloa polystachya plants during the removal of a pyrethroid-based pesticide. Plants were inoculated or not with the Trichoderma consortium and exposed to commercial pesticide H24®, based on pyrethroids. Pesticide application resulted in significant reduction in root protein content (58%), but enhanced content of malondialdehyde (MDA) in shoots, superoxide dismutase (SOD) activity in shoots and roots, and catalase (CAT) activity in roots. Inoculation of Trichoderma consortium in E. polystachya exposed to the pesticide resulted in increased protein content in roots and MDA content in shoots (2-fold). Trichoderma consortium improved protein content and SOD activity (140-fold) in plants. Fungal inoculation increased the removal (97.9%) of the pesticide in comparison to the sole effect of plants (33.9%). Results allow further understanding about the responses of the interaction between plants and root-associated fungi to improving the assisted-phytoremediation of solid matrices contaminated with organic pesticides.

This original paper describes the positive role of the Trichoderma sp. consortium on favoring the removal of a pyretrhoid-based pesticide. This is one of first reports on analyzing the influence of a Trichoderma consortium on the oxidative stress and antioxidant response of Echinochloa polystachya in presence of the pesticide. This experimental approach provides a new alternative for further fungal assisted-phytoremediation of a pyretrhoid-based pesticide.

Case report: Renal adenoma in a captive ocelot (Leopardus pardalis) in Costa Rica.

Reports of renal neoplasia are rare in neotropical wildcats. Ocelots (Leopardus pardalis) are medium-sized wildcats living in America's tropical forests. A 12-year-old captive ocelot was diagnosed with a renal mass occupying approximately 25% of the total right kidney volume. The tissue was stained with routine hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS). Immunohistochemistry with the following markers was performed: cytokeratin (CK) AE1/AE3, CK19, CK 7, CD10, vimentin, Melan A, HMB45, Pax-8, and Wilms' tumor 1 (WT1). Histopathology revealed a well-differentiated epithelial tubular neoplasia with less than one mitotic figure per 2.37mm2 field. Vimentin and Pax-8 were the only positive markers. Immunohistochemically, neoplasia was diagnosed as a renal adenoma. Renal adenomas are seldom reported in neotropical wildcats. Reports on wild species are valuable for properly establishing a clinical prognosis for captive species. To the best of our knowledge, this is the first report that provides detailed microscopic and immunohistochemical descriptions of renal adenoma in a captive ocelot.

Molecular, morphological and histopathological evidence of Spirometra mansoni in wild and domestic animals from Costa Rica.

Spirometra mansoni is a diphyllobothroid cestode and one of the causing agents of sparganosis, a zoonotic foodborne and waterborne infection in humans. This parasite has an indirect life cycle with domestic and wild canids or felids as definitive hosts. The last report of S. mansoni in Costa Rica was done in 2004 by morphological assessment of worms, whereas molecular evidence of this species was obtained recently in the Americas. Herein, we present seven cases of spirometrosis in four dogs, three cats and a coyote from different regions of Costa Rica occurring in a time span of a year. Dog cases presented vomiting, hyporexia, lethargy and diarrhea, whereas cats were mostly asymptomatic. Moreover, the coyote was found with Spirometra sp. proglottids incidentally. Cytochrome oxidase subunit 1 (cox1) sequences of eggs or proglottids derived from all cases were analyzed with a Bayesian Inference phylogenetic tree and a haplotype network. These analyses showed the clustering of S. mansoni from Costa Rica with other sequences derived from Asia and America. Moreover, cox1 sequences clustered in two separate haplotypes, suggesting the high genetic diversity of the species. The present cases represent the first molecular evidence of the parasite in Central America; thus, extending its known range in the American continent.

Growth, tolerance, and enzyme activities of Trichoderma strains in culture media added with a pyrethroids-based insecticide.

The application of pyrethroids and carbamates represents an environmental risk and may exert adverse effects on beneficial microorganisms such as Trichoderma, which contribute to the biocontrol of several fungal phytopathogens. This research evaluated the tolerance of several strains of Trichoderma to a selected culture medium contaminated with a commercial insecticide (H24®) composed of pyrethroids, permethrin and prallethrin, and carbamate propoxur, and determined the influence of this insecticide on the release of enzymes such as chitinases, peroxidases, and endoglucanases by a consortium of selected Trichoderma strains grown in liquid culture medium. Four out of 10 Trichoderma strains showed tolerance to 200ppm (∼48.3% of growth) of the commercial insecticide after 96h of exposure to a contaminated solid medium. After eight days of growth in liquid culture, the insecticide enhanced extracellular protein content and peroxidase activities in the Trichoderma consortium but decreased both chitinase and glucanase activities. These fungal responses should be considered when implementing strategies that combine alternative pesticides and fungal biocontrollers for managing fungal phytopathogens.

Experimental infection of Artibeus lituratus bats and no detection of Zika virus in neotropical bats from French Guiana, Peru, and Costa Rica suggests a limited role of bats in Zika transmission.

Bats are important natural reservoir hosts of a diverse range of viruses that can be transmitted to humans and have been suggested to play an important role in the Zika virus (ZIKV) transmission cycle. However, the exact role of these animals as reservoirs for flaviviruses is still controversial. To further expand our understanding of the role of bats in the ZIKV transmission cycle in Latin America, we carried out an experimental infection in wild-caught Artibeus lituratus bats and sampled several free-living neotropical bats across three countries of the region. Experimental ZIKV infection was performed in wild-caught adult bats (4 females and 5 males). The most relevant findings were hemorrhages in the bladder, stomach and patagium. Significant histological findings included inflammatory infiltrate consisting of a predominance of neutrophils and lymphocytes, in addition to degeneration in the reproductive tract of males and females. This suggests that bat reproduction might be at some level affected by ZIKV. Leukopenia was also observed in some inoculated animals. Hemorrhages, genital alterations, and leukopenia are suggested to be caused by ZIKV; however, since these were wild-caught bats, we cannot exclude other agents. Detection of ZIKV by qPCR was observed at low concentrations in only two urine samples in two inoculated animals. All other animals and tissues tested were negative. Finally, no virus-neutralizing antibodies were found in any animal. To determine ZIKV infection in nature, the blood of a total of 2056 bats was sampled for ZIKV detection by qPCR. Most of the sampled individuals belonged to the genus Pteronotus sp. (23%), followed by the species Carollia sp. (17%), Anoura sp. (14%), and Molossus sp. (13.7%). No sample of any tested species was positive for ZIKV by qPCR. These results together suggest that bats are not efficient amplifiers or reservoirs of ZIKV and may not have an important role in ZIKV transmission dynamics.

Cellulase and chitinase activities and antagonism against Fusarium oxysporum f.sp. cubense race 1 of six Trichoderma strains isolated from Mexican maize cropping.

OBJECTIVE: To evaluate the enzymatic and biocontrol capacity of native Trichoderma strains isolated from corn crops in Irapuato (state of Guanajuato) and Napízaro (state of Michoacán), Mexico. RESULTS: Six native strains from Irapuato and Napízaro were tested, with five of them identified as T. harzianum and one as T. tomentosum. The six strains qualitatively and quantitatively showed enzyme activity for cellulase and chitinase. The best results were obtained for strains IrV6SIC7 and MichV6S2C2 with 878 IU L-1 of chitinase and 1323 IU L-1 of cellulase, respectively. All Trichoderma strains acted antagonistically toward Fusarium oxysporum f.sp. cubense race 1 (FocR1), with percentages of inhibition that ranged from 9 to 54%. In addition, the microscopic analysis allowed visualizing the mechanisms of mycoparasitism and antibiosis by either IrV6SIC7 or MichV6S2C2. The latter effects indicate that the tested native Trichoderma strains isolated from corn crops possessed enzymatic mechanisms as a strategy for biocontrolling FocR1 strains. CONCLUSION: The enzyme production by the Trichoderma strains represents a potential biotechnological utilization for either agricultural or industrial purposes.

Seasonal changes in the diversity, host preferences and infectivity of mosquitoes in two arbovirus-endemic regions of Costa Rica.

BACKGROUND: Mosquitoes are vectors of various arboviruses belonging to the genera Alphavirus and Flavivirus, and Costa Rica is endemic to several of them. The aim of this study was to describe and analyze the community structure of such vectors in Costa Rica. METHODS: Sampling was performed in two different coastal locations of Costa Rica with evidence of arboviral activity during rainy and dry seasons. Encephalitis vector surveillance traps, CDC female gravid traps and ovitraps were used. Detection of several arboviruses by Pan-Alpha and Pan-Flavi PCR was attempted. Blood meals were also identified. The Normalized Difference Vegetation Index (NDVI) was estimated for each area during the rainy and dry seasons. The Chao2 values for abundance and Shannon index for species diversity were also estimated. RESULTS: A total of 1802 adult mosquitoes belonging to 55 species were captured, among which Culex quinquefasciatus was the most caught species. The differences in NDVI were higher between seasons and between regions, yielding lower Chao-Sørensen similarity index values. Venezuelan equine encephalitis virus, West Nile virus and Madariaga virus were not detected at all, and dengue virus and Zika virus were detected in two separate Cx. quinquefasciatus specimens. The primary blood-meal sources were chickens (60%) and humans (27.5%). Both sampled areas were found to have different seasonal dynamics and population turnover, as reflected in the Chao2 species richness estimation values and Shannon diversity index. CONCLUSION: Seasonal patterns in mosquito community dynamics in coastal areas of Costa Rica have strong differences despite a geographical proximity. The NDVI influences mosquito diversity at the regional scale more than at the local scale. However, year-long continuous sampling is required to better understand local dynamics.

Parasitic fauna of bats from Costa Rica.

Bats are important reservoirs and spreaders of pathogens, including those of zoonotic concern. Though Costa Rica hosts one of the highest bat species' diversity, no information is available about their parasites. In order to investigate the occurrence of vector-borne pathogens (VBPs) and gastrointestinal (GI) parasites of chiropterans from this neotropical area, ectoparasites (n = 231) and stools (n = 64) were collected from 113 bats sampled in Santa Cruz (site 1) and Talamanca (site 2). Mites, fleas and ticks were morphologically and molecularly identified, as well as pathogens transmitted by vectors (VBPs, i.e., Borrelia spp., Rickettsia spp., Bartonella spp.) and from feces, such as Giardia spp., Cryptosporidium spp. and Eimeria spp. were molecularly investigated. Overall, 21 bat species belonging to 15 genera and 5 families were identified of which 42.5% were infested by ectoparasites, with a higher percentage of mites (38.9%, i.e., Cameronieta sp. and Mitonyssoides sp.) followed by flies (2.6%, i.e., Joblingia sp.) and tick larvae (1.7%, i.e., Ornithodoros sp.). Rickettsia spp. was identified in one immature tick and phylogenetically clustered with two Rickettsia species of the Spotted Fever Group (i.e., R. massiliae and R. rhipicephali). The frequency of GI parasite infection was 14%, being 3.1% of bats infected by Giardia spp. (un-identified non-duodenalis species), 1.5% by Eimeria spp. and 9.4% by Cryptosporidium spp. (bat and rodent genotypes; one C. parvum-related human genotype). The wide range of ectoparasites collected coupled with the detection of Rickettsia sp., Giardia and Cryptosporidium in bats from Costa Rica highlight the role these mammals may play as spreaders of pathogens and the need to further investigate the pathogenic potential of these parasites.

Phytoremediation of soil contaminated with weathered petroleum hydrocarbons by applying mineral fertilization, an anionic surfactant, or hydrocarbonoclastic bacteria.

This study evaluated the effect of the application of mineral fertilization (F), the anionic surfactant Triton X-100 (TX100), or the inoculation with a hydrocarbooclastic bacterial consortium (BCons) on the growth of Clitoria ternatea during the phytoremediation of a Gleysol contaminated with weathered petroleum hydrocarbons (39,000 mg kg-1 WPH) collected from La Venta, Tabasco (Mexico). The experiment consisted of a completely randomized design with seven treatments and four replications each under greenhouse conditions. The application of F (biostimulation) increased plant growth and biomass production; in contrast, TX100 only favored root biomass (11%) but significantly favored WPH degradation. Bioaugmentation with BCons did not show significant effects on plant growth. Nevertheless, the combination of biostimulation with bioaugmentation (BCons + F, BCons + TX100, and BCons + F+TX100) enhanced plant growth, hydrocarbonoclastic bacteria population, and WPH degradation when compared to treatments with the single application of bioaugmentation (BCons) or biostimulation (F).

Application of mineral fertilization and commercial surfactant favored root biomass and degradation of weathered petroleum hydrocarbons (WPH). The reintroduction of hydrocarbonoclastic and surfactant-producer bacteria did not enhance plant growth but significantly contributed on WPH degradation from a chronically contaminated soil.

Short-Term Evaluation of the Spatial Distribution of Trophic Groups of Amoebae in the Rhizosphere of Zea mays Inoculated with Rhizophagus intraradices.

Primary production in terrestrial ecosystems is sustained by plants, microbiota, and fungi, which are the major organic matter providers in the root zone, setting in motion the soil food webs. Predators like soil amoebae voraciously feed on bacteria, fungi, and microbial eukaryotes releasing the nutrients sequestered in their biomass. Early food web setting up is crucial for seedling nutrition and its further development after establishment. Mycorrhizal fungi are more than phosphorus providers, and we wonder what their role is in structuring the predators' trophic groups in the root zone. We evaluated the effect of Rhizophagus intraradices inoculated in Zea mays (mycorrhizosphere), on the structuration of amoebae trophic groups along vertical and horizontal (3, 6, and 9 cm) soil distribution when compared to un-inoculated plants, after 20 days in microcosms. Amoebae species richness was highest in non-mycorrhizal seedlings in the root zone at 6- to 9-cm depth, and 3 cm away from plants. More bacterial species are needed when plants are devoid of mycorrhiza, and their influence is constrained 3 cm away from roots. Higher diversity of trophic groups was recorded at mycorrhizal seedlings and at the compartment influenced by the mycelium at 6- to 9-cm depth. The highest bacterivorous diversity, higher number of rare species and protozoa-eating amoebae, and the absence of fungivorous group recorded at the mycorrhizosphere of Z. mays, indicate that the community was very different from the non-mycorrhizal plants. We conclude that the arbuscular mycorrhizal fungus exerts significant changes on the community of trophic groups of amoebae.

Boid Inclusion Body Disease Is Also a Disease of Wild Boa Constrictors.

Reptarenaviruses cause boid inclusion body disease (BIBD), a potentially fatal disease, occurring in captive constrictor snakes boas and pythons worldwide. Classical BIBD, characterized by the formation of pathognomonic cytoplasmic inclusion bodies (IBs), occurs mainly in boas, whereas in pythons, for example, reptarenavirus infection most often manifests as central nervous system signs with limited IB formation. The natural hosts of reptarenaviruses are unknown, although free-ranging/wild constrictor snakes are among the suspects. Here, we report BIBD with reptarenavirus infection in indigenous captive and wild boid snakes in Costa Rica using histology, immunohistology, transmission electron microscopy, and next-generation sequencing (NGS). The snakes studied represented diagnostic postmortem cases of captive and wild-caught snakes since 1989. The results from NGS on archival paraffin blocks confirm that reptarenaviruses were already present in wild boa constrictors in Costa Rica in the 1980s. Continuous sequences that were de novo assembled from the low-quality RNA obtained from paraffin-embedded tissue allowed the identification of a distinct pair of reptarenavirus S and L segments in all studied animals; in most cases, reference assembly could recover almost complete segments. Sampling of three prospective cases in 2018 allowed an examination of fresh blood or tissues and resulted in the identification of additional reptarenavirus segments and hartmanivirus coinfection. Our results show that BIBD is not only a disease of captive snakes but also occurs in indigenous wild constrictor snakes in Costa Rica, suggesting boa constrictors to play a role in natural reptarenavirus circulation. IMPORTANCE The literature describes cases of boid inclusion body disease (BIBD) in captive snakes since the 1970s, and in the 2010s, others and ourselves identified reptarenaviruses as the causative agent. BIBD affects captive snakes globally, but the origin and the natural host of reptarenaviruses remain unknown. In this report, we show BIBD and reptarenavirus infections in two native Costa Rican constrictor snake species, and by studying archival samples, we show that both the viruses and the disease have been present in free-ranging/wild snakes in Costa Rica at least since the 1980s. The diagnosis of BIBD in wild boa constrictors suggests that this species plays a role in the circulation of reptarenaviruses. Additional sample collection and analysis would help to clarify this role further and the possibility of, e.g., vector transmission from an arthropod host.

Passive epidemiological surveillance in wildlife in Costa Rica identifies pathogens of zoonotic and conservation importance.

Epidemiological surveillance systems for pathogens in wild species have been proposed as a preventive measure for epidemic events. These systems can minimize the detrimental effects of an outbreak, but most importantly, passive surveillance systems are the best adapted to countries with limited resources. Therefore, this research aimed to evaluate the technical and infrastructural feasibility of establishing this type of scheme in Costa Rica by implementing a pilot program targeting the detection of pathogens of zoonotic and conservation importance in wildlife. Between 2018 and 2020, 85 carcasses of free-ranging vertebrates were admitted for post-mortem and microbiology analysis. However, we encountered obstacles mainly related to the initial identification of cases and limited local logistics capacity. Nevertheless, this epidemiological surveillance scheme allowed us to estimate the general state of health of the country's wildlife by establishing the causes of death according to pathological findings. For instance, 60% (51/85) of the deaths were not directly associated with an infectious agent. Though in 37.6% (32/85) of these cases an infectious agent associated or not with disease was detected. In 27.1% (23/85) of the cases, death was directly related to infectious agents. Furthermore, 12.9% (11/85), the cause of death was not determined. Likewise, this wildlife health monitoring program allowed the detection of relevant pathogens such as Canine Distemper Virus, Klebsiella pneumoniae, Angiostrongylus spp., Baylisascaris spp., among others. Our research demonstrated that this passive surveillance scheme is cost-effective and feasible in countries with limited resources. This passive surveillance can be adapted to the infrastructure dedicated to monitoring diseases in productive animals according to the scope and objectives of monitoring wildlife specific to each region. The information generated from the experience of the initial establishment of a WHMP is critical to meeting the challenges involved in developing this type of scheme in regions with limited resources and established as hotspots for emerging infectious diseases.

Platelet depletion does not alter the course of Brucella abortus infection in vivo.

Brucellosis is a bacterial disease of animals and a zoonotic infection. Thrombocytopenia is a common outcome in long-lasting brucellosis in humans. Likewise, ex vivo experiments have shown that platelets may play a role in Brucella abortus infections. Following these reports, we explored the course of brucellosis in thrombocytopenic mice, using the non-toxic low-molecular-weight aspercetin protein that depletes platelets in vivo. Aspercetin does not induce systemic hemorrhage or inflammation, and when injected into mice, it generates a rapid dose-dependent drop in platelet counts without affecting central organs, disrupting hematological parameters, or the proinflammatory cytokine profile. Compared to the B. abortus infected control group, the infected thrombocytopenic mice did not show significant differences in the hematological profiles, pathological score, spleen, liver histopathology, or bacterial loads. Except for IL-6, which was higher in the infected thrombocytopenic mice, the TNF-α, IFN-γ and IL-10 did not significantly differ with the PBS-infected group. The results indicate that platelets do not play a significant role in modulating Brucella infection in vivo at the early stages of infection, which is commensurate with the stealthy strategy followed by Brucella organisms at the onset of the disease.

Serological Positivity against Selected Flaviviruses and Alphaviruses in Free-Ranging Bats and Birds from Costa Rica Evidence Exposure to Arboviruses Seldom Reported Locally in Humans.

Arboviruses have two ecological transmission cycles: sylvatic and urban. For some, the sylvatic cycle has not been thoroughly described in America. To study the role of wildlife in a putative sylvatic cycle, we sampled free-ranging bats and birds in two arbovirus endemic locations and analyzed them using molecular, serological, and histological methods. No current infection was detected, and no significant arbovirus-associated histological changes were observed. Neutralizing antibodies were detected against selected arboviruses. In bats, positivity in 34.95% for DENV-1, 16.26% for DENV-2, 5.69% for DENV-3, 4.87% for DENV-4, 2.43% for WNV, 4.87% for SLEV, 0.81% for YFV, 7.31% for EEEV, and 0.81% for VEEV was found. Antibodies against ZIKV were not detected. In birds, PRNT results were positive against WNV in 0.80%, SLEV in 5.64%, EEEV in 8.4%, and VEEV in 5.63%. An additional retrospective PRNT analysis was performed using bat samples from three additional DENV endemic sites resulting in a 3.27% prevalence for WNV and 1.63% for SLEV. Interestingly, one sample resulted unequivocally WNV positive confirmed by serum titration. These results suggest that free-ranging bats and birds are exposed to not currently reported hyperendemic-human infecting Flavivirus and Alphavirus; however, their role as reservoirs or hosts is still undetermined.

Effect of Predation by Colpoda sp. in Nitrogen Fixation Rate of Two Free-Living Bacteria.

Biological nitrogen fixation is limited to several groups of prokaryotes, some of them reduce nitrogen as free-living nitrogen-fixing bacteria. Protozoa predation on these latter releases sequestered nitrogen that may enhance the formation of new bacterial biomass and possibly increase nitrogen fixation within soil microbial communities. We aim to evaluate the predation effect of Colpoda sp. on two nitrogen fixers: Azospirillum lipoferum and Stenotrophomonas sp. during their lag, early exponential, and exponential phases. The kinetics of bacterial population growth was determined in the predators' presence or absence and the effect of predation on the rate of N fixation was evaluated through the reduction of acetylene to ethylene technique. Colpoda sp. showed a non-significant difference in preferences between the two species offered as prey. Consequently, the abundance of A. lipoferum and Stenotrophomonas sp. decreased significantly due to predator's pressure and both species responded by increasing their specific growth rate. Likewise, predation promoted greater nitrogen fixation rate by CFU during the lag phase in A. lipoferum (0.20 nM/CFU with predation vs 0.09 nM/CFU without predation) and Stenotrophomonas sp. (0.22 nM/CFU vs 0.09 nM/CFU respectively). During early exponential phase (29 h), the rate diminished to 0.13 and 0.05 nM/CFU in A. lipoferum and to 0.09 nM/CFU and 0.05 nM/CFU in Stenotrophomonas sp. Finally, during the exponential phase (52 h), only A. lipoferum without predation produced 0.003 nM/CFU of ethylene. Thus, the nitrogenase activity was higher in the lag and the early exponential phases when predator activity was involved.

Serosurvey in Two Dengue Hyperendemic Areas of Costa Rica Evidence Active Circulation of WNV and SLEV in Peri-Domestic and Domestic Animals and in Humans.

Costa Rica harbors several flaviviruses, including Dengue (DENV), Zika (ZIKV), West Nile virus (WNV), and Saint Louis encephalitis virus (SLEV). While DENV and ZIKV are hyperendemic, previous research indicates restricted circulation of SLEV and WNV in animals. SLEV and WNV seroprevalence and high transmission areas have not yet been measured. To determine the extents of putative WNV and SLEV circulation, we sampled peri-domestic and domestic animals, humans, and mosquitoes in rural households located in two DENV and ZIKV hyperendemic regions during the rainy and dry seasons of 2017-2018 and conducted plaque reduction neutralization test assay for serology (PRNT) and RT-PCR for virus detection. In Cuajiniquil, serological evidence of WNV and SLEV was found in equines, humans, chickens, and wild birds. Additionally, five seroconversion events were recorded for WNV (2 equines), SLEV (1 human), and DENV-1 (2 humans). In Talamanca, WNV was not found, but serological evidence of SLEV circulation was recorded in equines, humans, and wild birds. Even though no active viral infection was detected, the seroconversion events recorded here indicate recent circulation of SLEV and WNV in these two regions. This study thus provides clear-cut evidence for WNV and SLEV presence in these areas, and therefore, they should be considered in arboviruses differential diagnostics and future infection prevention campaigns.

Esophageal spirocercosis with pulmonary egg deposition and secondary hypertrophic osteopathy in a dog from Costa Rica.

Spirocerca lupi is the etiologic agent of spirocercosis in canids from the tropical and subtropical regions of the world. A 1-year-old dog was presented with weight loss and diarrhea. Upon examination, thickened limbs, a mass in the lung's right middle lobe and an esophageal mass were found. The pulmonary mass aspirate revealed S. lupi eggs surrounded by a purulent infiltrate. Limb bone biopsies showed irregular mineralization and cartilaginous metaplasia. Treatment with doramectin resolved the initial clinical signs. However, regurgitation was reported 5 months later. Therefore, the dog received additional doramectin and later vomited a worm morphologically consistent with a S. lupi adult. Herein we report the esophageal migration of S. lupi with egg deposition to the lung parenchyma, accompanied by secondary hypertrophic osteopathy. This report highlights the presence of S. lupi in Costa Rica and the importance of including this parasite as a differential diagnosis when compatible signs are identified.

Growth and lead uptake by Parkinsonia aculeata L. inoculated with Rhizophagus intraradices.

The increased lead (Pb) pollution in the biosphere has resulted in serious environmental problems, so it is essential to evaluate phytoremediation strategies for contaminated soils. This study evaluated the growth and Pd absorption capacity of Pakinsonia aculeata, inoculated with an arbuscular mycorrhizal fungus (Rhizophagus intraradices) over 18 weeks under greenhouse conditions. Treatments included inoculated and non-inoculated plants combined with six Pb concentrations (0, 40, 80, 160, 320, 640 mg·L-1) in the form of Pb(NO3)2. Results showed that mycorrhizal colonization in inoculated plants ranged from 5.0 to 6.7% and favored plant growth. Pb levels and AMF-inoculation had no effects on chlorophyll fluorescence values. AMF-plants absorbed significantly more Pb in roots (237.97 mg·kg-1) than control plants (202.85 mg·kg-1), as well as high translocation to shoots (27.02 mg·kg-1) under the high Pb dose. The increase in Pb concentration reduced the P concentration in roots, and the P and N concentrations in shoots; however, the absorption and translocation of Ca and Mg was increased in shoots. Inoculation of R. intraradices improved both growth and Pb uptake of P. aculeata, under greenhouse conditions suggesting that this tree species may be potentially studied for detoxifying Pb-polluted soils.

Experiences and meanings from collective occupations: a look at the well-being/psychosocial discomfort expressed by conducted public transportation drivers / Experiências e significações de ocupações coletivas: um olhar para o bem-estar psicossocial/desconforto expresso por condutores de transporte público / Experiencias y significados desde las ocupaciones colectivas: una mirada al bienestar/malestar psicosocial expresado por conductores del transporte público

Abstract The article describes experiences from collective occupations in the territory and the meaning given to the psychosocial well-being/discomfort by drivers of the public transport tendered in the province of Concepción, Chile. In this research, the methodology was qualitative in nature, with a phenomenological approach. The information was collected through the application of seventeen semi-structured interviews with bus line drivers, which were later transcribed. The texts of these transcripts were the material for the categorical analysis, which allowed defining themes, categories, encodings, and analysis subcategories. Among the most relevant results, the social construction of the figure that is currently perceived as the driver of the public transport tendered in Concepción stands out, understood as a collective occupation in movement and which is dependent on the unique characteristics of the territory. The Conclusions in this study allow us to observe that drivers work long hours in exchange for the economic benefits that work grants them, in some cases showing occupational alienation. Different territories are identified; the terminal as a meeting space where "we are all friends", a relationship that changes when we go out onto the street, a space that drivers describe as the "jungle". Thus, the street is identified as a field of struggle, competition, and permanent conflict. It is possible to show that the union organization of workers contributes to the provision of spaces in the terminal territory, allowing collective occupation.

Resumo O artigo exposto descreve as experiências das ocupações coletivas no território e o significado atribuído ao bem-estar/desconforto psicossocial dos motoristas de transporte público ofertados na província de Concepción, Chile. Nesta pesquisa, a metodologia utilizada foi qualitativa, com abordagem fenomenológica. As informações. Foram coletadas por meio da aplicação de dezessete entrevistas semiestruturadas com motoristas de linhas de ônibus, que foram posteriormente transcritas. Os textos destas transcrições foram o material para a análise categórica, o que permitiu definir temas, categorias, codificações e subcategorias de análise. Entre os resultados mais relevantes, destaca-se a construção social da figura que atualmente é percebida como propulsora do sistema de transporte público em Concepción, Chile, entendida como ocupação coletiva em movimento e dependente das características únicas do território. As conclusões deste estudo permitem observar que os motoristas trabalham longas horas em troca dos benefícios econômicos que o trabalho lhes concede, em alguns casos apresentando alienação ocupacional. Diferentes territórios foram identificados; sendo o terminal de ônibus um espaço de encontro onde "somos todos amigos", sendo que a relação muda quando saímos para a rua, um espaço que os motoristas descrevem como "selva". Assim, a rua é identificada como um campo de luta, competição e conflito permanente. É possível evidenciar que a organização sindical dos trabalhadores contribui para a disponibilização de espaços no território do terminal, permitindo a ocupação coletiva.

Resumen El artículo que se expone describe experiencias desde las ocupaciones colectivas en el territorio y el significado otorgado al bienestar/malestar psicosocial por conductores del transporte público licitado de la provincia de Concepción, Chile. En esta investigación, la metodología empleada fue de carácter cualitativo, con enfoque fenomenológico. La información fue recolectada mediante la aplicación de diecisiete entrevistas semiestructuradas a conductores de líneas de buses, las que posteriormente fueron transcritas. Los textos de estas transcripciones fueron el material para el análisis categorial, que permitió definir temas, categorías, codificaciones y subcategorías de análisis. Entre los resultados más relevantes se destaca la construcción social de la figura que actualmente se percibe del conductor del transporte público licitado de Concepción, entendida como ocupación colectiva en movimiento y que es dependiente de las características singulares del territorio. Las Conclusiones en este estudio, permiten observar que los conductores realizan extensas jornadas laborales a cambio de los beneficios económicos que el trabajo les otorga, dejando en evidencia en algunos casos una alienación ocupacional. Se identifican diferentes territorios; el terminal como un espacio de encuentro en donde "todos somos amigos", relación que cambia al salir al territorio de la calle, espacio que los conductores describen como la "selva". Así, la calle es identificada como un campo de lucha, competencia y conflicto permanente. Es posible evidenciar que la organización sindical de los trabajadores contribuye a la disposición de espacios en el territorio terminal, permitiendo la ocupación colectiva.

In vivo animal models confirm an increased virulence potential and pathogenicity of the NAP1/RT027/ST01 genotype within the Clostridium difficile MLST Clade 2.

BACKGROUND: Based on MLST analyses the global population of C. difficile is distributed in eight clades, of which Clade 2 includes the "hypervirulent" NAP1/RT027/ST01 strain along with various unexplored sequence types (STs). METHODS: To clarify whether this clinically relevant phenotype is a widespread feature of C. difficile Clade 2, we used the murine ileal loop model to compare the in vivo pro-inflammatory (TNF-α, IL-1ß, IL-6) and oxidative stress activities (MPO) of five Clade 2 clinical C. difficile isolates from sequence types (STs) 01, 41, 67, and 252. Besides, we infected Golden Syrian hamsters with spores from these strains to determine their lethality, and obtain a histological evaluation of tissue damage, WBC counts, and serum injury biomarkers (LDH, ALT, AST, albumin, BUN, creatinine, Na+, and Cl-). Genomic distances were calculated using Mash and FastANI to explore whether the responses were dictated by phylogeny. RESULTS: The ST01 isolate tested ranked first in all assays, as it induced the highest overall levels of pro-inflammatory cytokines, MPO activity, epithelial damage, biochemical markers, and mortality measured in both animal models. Statistically indistinguishable or rather similar outputs were obtained for a ST67 isolate in tests such as tissue damage, neutrophils count, and lethal activity. The results recorded for the two ST41 isolates tested were of intermediate magnitude and the ST252 isolate displayed the lowest pathogenic potential in all animal experiments. This ordering matched the genomic distance of the ST01 isolate to the non-ST01 isolates. CONCLUSIONS: Despite their close phylogenic relatedness, our results demonstrate differences in pathogenicity and virulence levels in Clade 2 C. difficile strains, confirm the high severity of infections caused by the NAP1/RT027/ST01 strain, and highlight the importance of C. difficile typing.