Techno-economic assessment of biopolymer production from methane and volatile fatty acids: effect of the reactor size and biomass concentration on the poly(3-hydroxybutyrate-co-3-hydroxyvalerate) selling price.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) is a biobased and biodegradable polymer that could efficiently replace fossil-based plastics. However, its widespread deployment is slowed down by the high production cost. In this work, the techno-economic assessment of the process for producing poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from low-cost substrates, such as methane and valeric acid derived from the anaerobic digestion of organic wastes, is proposed. Several strategies for cost abatement, such as the use of a mixed consortium and a line for reagent recycling during downstream, were adopted. Different scenarios in terms of production, from 100 to 100,000 t/y, were analysed, and, for each case, the effect of the reactor volume (small, medium and large size) on the selling price was assessed. In addition, the effect of biomass concentration was also considered. Results show that the selling price of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) is minimum for a production plant with 100,000 t/y capacity, accounting for 18.4 €/kg, and highly influenced by the biomass concentration since it can be reduced up to 8.6 €/kg by increasing the total suspended solids from 5 to 30 g/L, This adjustment aligns the breakeven point of PHBV with the reported average commercial price.

Polyhydroxyalkanoate recovery overview: properties, characterizations, and extraction strategies.

Due to their excellent properties, polyhydroxyalkanoates are gaining increasing recognition in the biodegradable polymer market. These biogenic polyesters are characterized by high biodegradability in multiple environments, overcoming the limitation of composting plants only and their versatility in production. The most consolidated techniques in the literature or the reference legislation for the physical, chemical and mechanical characterisation of the final product are reported since its usability on the market is still linked to its quality, including the biodegradability certificate. This versatility makes polyhydroxyalkanoates a promising prospect with the potential to replace fossil-based thermoplastics sustainably. This review analyses and compares the physical, chemical and mechanical properties of poly-ß-hydroxybutyrate and poly-ß-hydroxybutyrate-co-ß-hydroxyvalerate, indicating their current limitations and strengths. In particular, the copolymer is characterised by better performance in terms of crystallinity, hardness and workability. However, the knowledge in this area is still in its infancy, and the selling prices are too high (9-18 $ kg-1). An analysis of the main extraction techniques, established and in development, is also included. Solvent extraction is currently the most widely used method due to its efficiency and final product quality. In this context, the extraction phase of the biopolymer production process remains a major challenge due to its high costs and the need to use non-halogenated toxic solvents to improve the production of good-quality bioplastics. The review also discusses all fundamental parameters for optimising the process, such as solubility and temperature.

Production of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from methane and volatile fatty acids: properties, metabolic routes and current trend.

Polyhydroxyalkanoates (PHAs) are biobased and biodegradable polymers that could effectively replace fossil-based and non-biodegradable plastics. However, their production is currently limited by the high production costs, mainly due to the costly carbon sources used, low productivity and quality of the materials produced. A potential solution lies in utilizing cheap and renewable carbon sources as the primary feedstock during the biological production of PHAs, paving the way for a completely sustainable and economically viable process. In this review, the opportunities and challenges related to the production of polyhydroxyalkanoates using methane and volatile fatty acids (VFAs) as substrates were explored, with a focus on poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate). The discussion reports the current knowledge about promising Type II methanotrophs, the impact of process parameters such as limiting nutrients, CH4:O2 ratio and temperature, the type of co-substrate and its concentration. Additionally, the strategies developed until now to enhance PHA production yields were also discussed.

Exploring New Strategies for Optimizing the Production of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from Methane and VFAs in Synthetic Cocultures and Mixed Methanotrophic Consortia.

In this work, the potential of a synthetic coculture and a mixed methanotrophic consortium to synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) from renewable and waste-based feedstocks was assessed batchwise. Methylocystis parvuscocultivated with Rhodococcus opacus and a Methylocystis-enriched culture previously grown on methane were subjected to nutrient starvation in a medium enriched with valeric acid (30% w w-1 of Ctot) or with a VFAs mixture containing acetic, propionic, butyric, and valeric acids (15% w w-1 of Ctot) under a CH4:O2 or air atmosphere. For all test series, pH was adjusted to 7 after adding the cosubstrates, and a negligible substrate consumption or polymer production was considered the end point of the trial. Results showed that valeric acid promoted PHBV accumulation in both cultures regardless of the atmosphere. Interestingly, the mixture of VFAs supported PHBV accumulation only in the presence of methane. The highest PHBV contents for the coculture and the mixed consortium, equal to 73.7 ± 2.5% w w-1 and 49.6 ± 13% w w-1, respectively, were obtained with methane and the VFAs mixture. This study demonstrates the suitability of cocultures and biobased cosubstrates for the sustainable production of the biodegradable polymer PHBV.

The co-conversion of methane and mixtures of volatile fatty acids into poly(3-hydroxybutyrate-co-3-hydroxyvalerate) expands the potential of an integrated biorefinery.

In this work, the potential of Methylocystis hirsuta to simultaneously use methane and volatile fatty acids mixtures for triggering PHBV accumulation was assessed for the first time batchwise. Biotic controls carried out with CH4 alone confirmed the inability of Methylocystis hirsuta to produce PHBV and achieved 71.2 ± 7 g m-3d-1 of PHB. Pure valeric acid and two synthetic mixtures simulating VFAs effluents from the anaerobic digestion of food waste at 35 °C (M1) and 55 °C (M2) were supplied to promote 3-HV inclusion. Results showed that pure valeric acid supported the highest polymer yields of 105.8 ± 9 g m-3d-1 (3-HB:3-HV=70:30). M1 mixtures led to a maximum of 103 ± 4 g m-3d-1 of PHBV (3-HB:3-HV=85:15), while M2 mixtures, which did not include valeric acid, showed no PHV synthesis. This suggested that the synthesis of PHBV from VFAs effluents depends on the composition of the mixtures, which can be tuned during the anaerobic digestion process.

Sustainable Process for the Production of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from Renewable Resources: A Simulation Study.

Bacterially produced polyhydroxyalkanoates are valuable substitutes for petrochemical plastics, but their current production capacities are very scarce. Producing poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-co-HV) from methane and odd-chain carbon fatty acids could make the production of this biodegradable polymer cost-effective. This study analyzes the main factors affecting methanotrophic growth and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) accumulation, simulating a pilot-scale process based on a double-stage approach. The effects of the nitrogen source and the oxygen partial pressure during a 20 day growth phase were studied; the cosubstrate concentration, the culture selected, and the methane partial pressure were investigated during the poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production stage performed within 15 days under nutrient starvation. Methylocystis parvus OBBP and Methylosinus thricosporum OB3b reached the maximum growth productivities with ammonium as the nitrogen source and oxygen at high partial pressure. The simulation of the PHB-co-HV accumulation revealed that methanotrophs could better accumulate the polymer with low valeric acid concentrations. A methane-abundant gas stream (0.5 atm of methane) could increase process yields up to 0.32 kg m-3 d-1.

Poly(3-hydroxybutyrate) production from methane in bubble column bioreactors: Process simulation and design optimization.

The pressing problem posed by plastic pollution has led to other, environmentally friendly alternatives, such as polyhydroxyalkanoates. This work proposes an innovative process to produce poly(3-hydroxybutyrate) by replacing expensive substrates, such as sugars, with methane. A two-step process was simulated: a first fermentation is performed in a continuous mode for 20 days to grow a strain belonging to the genus Methylocystis, while a second semi-continuous and nitrogen-limited fermentation is employed to induce the poly(3-hydroxybutyrate) accumulation within 12 days. The effects of the superficial gas velocity on the mass transfer and the poly(3-hydroxybutyrate) production yields were evaluated. Several scenarios were analyzed to optimize the geometry of the reactors and the methane utilization. The working volume of the reactors, as well as the presence of gas recycling stream, were shown to affect the global yields positively, while improving the aspect ratio from 8 to 19, with equal volume, lowered the fraction of poly(3-hydroxybutyrate) into the biomass by up to 37.5%.

Deep Neck Infections: Decisional Algorithm for Patients with Multiple Spaces Involvement.

BACKGROUND: Deep Neck Infections (DNIs) spread along fascial planes and involve neck spaces. Recently, their incidence has decreased due to the introduction of antibiotics; nevertheless, complications related to DNIs are often life-threatening. OBJECTIVE: The purpose of this article is focused on the identification of predisposing factors of these complications, as well as on the development of a reliable therapeutic algorithm. METHODS: Sixty patients with DNIs were enrolled from 2006 to 2019 for a retrospective study. The exclusion criteria for the present study were cellulitis, small abscesses responding to empiric or specific antibiotic therapy, or involvement of only one deep neck space. During the analysis, the following parameters of interest have been evaluated: gender, age, site of origin, pathways of spread, comorbidities, clinical features, bacteriology data, type of surgical approach required, complications, duration of hospitalization and mortality rate. On admission, microbial swab analysis was performed. RESULTS: Diabetes Mellitus (DM), Chronic Obstructive Pulmonary Disease (COPD), iron deficiency anemia and the involvement of multiple spaces have been associated with a significantly higher risk of developing complications. Most of our patients had polymicrobial infections. All patients underwent surgical drainage. The complication rate had occurred in 56.6% of patients, while death in 18.3%. CONCLUSION: DNIs represent a medical and surgical emergency with potentially serious complications; thus, avoidance of diagnostic delay is mandatory. Our preliminary data suggest the importance of evaluating the extent of infections because the involvement of multiple spaces requires timely surgery due to the higher risk of complications and mortality.

Covid-19 laryngectomized patients care, on field experience, and considerations.

Laryngectomized patients showed an unconventional response to SARS-CoV-2 viral infection. Here, we describe five different patient cases along with our interpretation of the phenomena and suggestions for their safe management.

Definition of miRNA Signatures of Nodal Metastasis in LCa: miR-449a Targets Notch Genes and Suppresses Cell Migration and Invasion.

Laryngeal cancer (LCa), a neoplasm of the head and neck region, is a leading cause of death worldwide. Surgical intervention remains the mainstay of LCa treatment, but a crucial point is represented by the possible nodal involvement. Therefore, it is urgently needed to develop biomarkers and therapeutic tools able to drive treatment approaches for LCa. In this study, we investigated deregulated microRNAs (miRNAs) in tissues from LCa patients with either lymph node metastases (N+) or not (N-). miRNA expression profiling was performed by a comprehensive PCR array and subsequent validation by RT-qPCR. Results showed a significant decrease of miR-449a expression in N+ compared to N- patients, and miR-133b down-modulation in LCa tissues compared to paired normal ones. Receiver operating characteristic (ROC) curve analysis revealed the potential diagnostic power of miR-133b for LCa detection. According to the validation results, we selected miR-449a for further in vitro studies. Ectopic miR-449a expression in the LCa cell line Hep-2 inhibited invasion and motility in vitro, slowed cell proliferation, and induced the downregulation of Notch1 and Notch2 as direct targets of miR-449a. Collectively, this study provides new promising biomarkers for LCa diagnosis and a new opportunity to use miR-449a for the treatment of nodal metastases in LCa patients.

Adenoid cystic carcinoma of the larynx in a 70-year-old patient: A case report.

Adenoid cystic carcinoma (ACC) is a relatively rare tumor that accounts for <1% of all head and neck malignancies. Laryngeal localization of ACC, which is most commonly hypoglottic, is relatively rare, occurring in 0.07-0.25% of all laryngeal tumors. ACC is characterized as a slow-growing tumor with a high recurrence rate, which often causes dyspnea and hoarseness. ACC exhibits a propensity for perineural invasion and thus, patients may experience pain as a late symptom of the disease. Distant metastasis occurs in 35-50% of cases and the lungs are the most common site of metastasis. Tumors are usually diagnosed by physical examination with fiberoscopy and computed tomography of the neck and chest, due to the high rate of lung metastases. The standard therapy for ACC is surgery followed by radiotherapy. In this study, a 70-year-old patient presented with laryngeal ACC, who underwent total laryngectomy with bilateral neck dissection and adjuvant radiotherapy, is presented. Follow-up examination performed 2 years after surgery revealed no evidence of locoregional recurrence or distant metastases. Previously published literature regarding ACC of the larynx was also reviewed.

Aggressiveness pattern and second primary tumor risk associated with basaloid squamous cell carcinoma of the larynx.

Basaloid squamous cell carcinoma (BSCC) is a rare, aggressive and distinct variant of squamous cell carcinoma (SCC) of the upper respiratory and digestive tract. We have evaluated disease specific survival (DSS) and overall survival (OS) through Kaplan-Meier method and mortality risk through univariate statistical analysis of Cox in 42 cases of BSCC and other 42 of laryngeal SCC (LSCC) matched for both age and sex. We demonstrated that laryngeal BSCC is a more aggressive tumor than LSCC as is associated to higher nodal recurrence of pathology (5 vs 2 patients, overall risk, OR 2.7), a reduced survival (median survival 34 vs 40 months, OR 3.2 for mortality); in addition, basaloid patients have a higher risk to be affected by second primary tumors (13 vs 3 patients, OR 5.8) and a higher probability to die for this second tumor (Hazard Risk, HR 4.4). The analysis of survival shows an increased mortality risk concurrent with the parameters assessed by univariate analyses that assume a predictive and statistical significance in second tumor and grading in basaloid LSSC.

Effect of liposomes, vitamins A and E nasal spray in reducing the healing time after septoplasty with concurrent turbinate surgery in adolescents.

BACKGROUND: This study evaluates endoscopic and quality of life (QoL) outcomes in two groups, composed by both allergic (A) and non-allergic (N-A) adolescents undergoing septoplasty with concurrent turbinate surgery by radiofrequency treated after surgery with nasal spray containing liposomes and vitamins A and E. METHODS: This double-blind randomized study was carried out on 40 patients (22 males, 18 females, mean age 15.5±1.0 years) undergoing septoplasty and volumetric tissue reduction of inferior turbinates by radiofrequency for post-traumatic deformities and severe nasal obstruction. We enrolled 20 subjects with positive skin prick tests and 20 with negative skin prick tests (SPT) to aeroallergens. All subjects underwent the Visual Analogue Scale (VAS) questionnaire and the nasal endoscopy score by Lund and Kennedy (LK) at baseline before surgery (T0) and 15 days (T15) and 21 days (T21) after treatment. After surgery, the nose was not packed and the day after surgery all subjects were randomized into two groups, experimental arm group I (GI) and control arm group II (GII) and given the treatment, nasal spray containing liposomes composed of phospholipids, fatty acids, and vitamin A and E for group I and saline solution (sodium chloride 0.9%) for group II. RESULTS: After therapy, the VAS and LK scores were better (P<0.05) in the investigational than in the control arm in both sub-groups (A and N-A). CONCLUSIONS: Our findings suggested that liposomal, vitamins A and E nasal spray improves the QoL in the early postoperative period, both in A and N-A sub-groups.

Orally-transmitted Chagas disease: Epidemiological, clinical, serological and molecular outcomes of a school microepidemic in Chichiriviche de la Costa, Venezuela.

Oral transmission of Trypanosoma cruzi is a frequent cause of acute Chagas disease (ChD). In the present cross-sectional study, we report the epidemiological, clinical, serological and molecular outcomes of the second largest outbreak of oral ChD described in the literature. It occurred in March 2009 in Chichiriviche de la Costa, a rural seashore community at the central littoral in Venezuela. The vehicle was an artisanal guava juice prepared at the local school and Panstrongylus geniculatus was the vector involved. TcI genotype was isolated from patients and vector; some showed a mixture of haplotypes. Using molecular markers, parasitic loads were high. Eighty-nine cases were diagnosed, the majority (87.5%) in school children 6-15 years of age. Frequency of symptomatic patients was high (89.9%) with long-standing fever in 87.5%; 82.3% had pericardial effusion detected by echocardiogram and 41% had EKG abnormalities. Three children, a pregnant woman and her stillborn child died (5.6% mortality). The community was addressed by simultaneous determination of specific IgG and IgM, confirmed with indirect hemagglutination and lytic antibodies. Determination of IgG and IgA in saliva had low sensitivity. No individual parasitological or serological technique diagnosed 100% of cases. Culture and PCR detected T. cruzi in 95.5% of examined individuals. Based on the increasing incidence of oral acute cases of ChD, it appears that food is becoming one of the most important modes of transmission in the Amazon, Caribbean and Andes regions of America.

Speech discrimination is impaired in parkinsonian patients: Expanding the audiologic findings of Parkinson's disease.

BACKGROUND AND OBJECTIVES: Hearing impairment (HI) has been previously demonstrated in patients with Parkinson's disease (PD). Pure Tone Audiometry (PTA) gives no information about patients' ability to hear and understand speech. To find out hearing ability and speech discrimination of PD patients, we expanded audiological evaluation by means of speech audiometry (SA). PATIENTS AND METHODS: We screened a series of consecutive PD patients. Severity of motor symptoms and staging were measured by the UPDRS-III and the H&Y scales. Audiometric evaluation consisted of a standardized audiological examination, PTA and SA. Healthy age- and sex-matched subjects were selected as controls. RESULTS: 45 PD patients and 45 healthy controls were enrolled. PTA confirmed our previous finding of high-frequency HI in PD patients. The mean values for the Speech Recognition Threshold were higher in PD patients as compared with controls. PD patients were more likely to have impaired speech discrimination profiles and higher disease stages. Neither the patients nor the controls showed a significant speech-tone dissociation and rollover phenomenon. CONCLUSION: Our results confirmed sensorineural HI in PD patients. Moreover, SA showed impaired speech discrimination abilities in PD patients as compared with control group thus expanding the audiologic findings of PD.

Temporal bone meningioma involving the middle ear: A case report.

Meningioma is a common intracranial tumor involving the meninges. The localization of this type of tumor is rarely extracranial due to its typically low invasive properties. Furthermore, invasion of the middle ear is exceptional. The present study reported a case of meningioma extending into the middle ear from the middle cranial fossa through the tegmen tympani. The clinical and pathological characteristics, as well as the outcome of the patient, were described.

Analytical performance of a multiplex Real-Time PCR assay using TaqMan probes for quantification of Trypanosoma cruzi satellite DNA in blood samples.

BACKGROUND: The analytical validation of sensitive, accurate and standardized Real-Time PCR methods for Trypanosoma cruzi quantification is crucial to provide a reliable laboratory tool for diagnosis of recent infections as well as for monitoring treatment efficacy. METHODS/PRINCIPAL FINDINGS: We have standardized and validated a multiplex Real-Time quantitative PCR assay (qPCR) based on TaqMan technology, aiming to quantify T. cruzi satellite DNA as well as an internal amplification control (IAC) in a single-tube reaction. IAC amplification allows rule out false negative PCR results due to inhibitory substances or loss of DNA during sample processing. The assay has a limit of detection (LOD) of 0.70 parasite equivalents/mL and a limit of quantification (LOQ) of 1.53 parasite equivalents/mL starting from non-boiled Guanidine EDTA blood spiked with T. cruzi CL-Brener stock. The method was evaluated with blood samples collected from Chagas disease patients experiencing different clinical stages and epidemiological scenarios: 1- Sixteen Venezuelan patients from an outbreak of oral transmission, 2- Sixty three Bolivian patients suffering chronic Chagas disease, 3- Thirty four Argentinean cases with chronic Chagas disease, 4- Twenty seven newborns to seropositive mothers, 5- A seronegative receptor who got infected after transplantation with a cadaveric kidney explanted from an infected subject. CONCLUSIONS/SIGNIFICANCE: The performing parameters of this assay encourage its application to early assessment of T. cruzi infection in cases in which serological methods are not informative, such as recent infections by oral contamination or congenital transmission or after transplantation with organs from seropositive donors, as well as for monitoring Chagas disease patients under etiological treatment.

The performance of laboratory tests in the management of a large outbreak of orally transmitted Chagas disease.

Orally transmitted Chagas disease (ChD), which is a well-known entity in the Brazilian Amazon Region, was first documented in Venezuela in December 2007, when 103 people attending an urban public school in Caracas became infected by ingesting juice that was contaminated with Trypanosoma cruzi. The infection occurred 45-50 days prior to the initiation of the sampling performed in the current study. Parasitological methods were used to diagnose the first nine symptomatic patients; T. cruzi was found in all of them. However, because this outbreak was managed as a sudden emergency during Christmas time, we needed to rapidly evaluate 1,000 people at risk, so we decided to use conventional serology to detect specific IgM and IgG antibodies via ELISA as well as indirect haemagglutination, which produced positive test results for 9.1%, 11.9% and 9.9% of the individuals tested, respectively. In other more restricted patient groups, polymerase chain reaction (PCR) provided more sensitive results (80.4%) than blood cultures (16.2%) and animal inoculations (11.6%). Although the classical diagnosis of acute ChD is mainly based on parasitological findings, highly sensitive and specific serological techniques can provide rapid results during large and severe outbreaks, as described herein. The use of these serological techniques allows prompt treatment of all individuals suspected of being infected, resulting in reduced rates of morbidity and mortality.

The performance of laboratory tests in the management of a large outbreak of orally transmitted Chagas disease

Orally transmitted Chagas disease (ChD), which is a well-known entity in the Brazilian Amazon Region, was first documented in Venezuela in December 2007, when 103 people attending an urban public school in Caracas became infected by ingesting juice that was contaminated with Trypanosoma cruzi. The infection occurred 45-50 days prior to the initiation of the sampling performed in the current study. Parasitological methods were used to diagnose the first nine symptomatic patients; T. cruzi was found in all of them. However, because this outbreak was managed as a sudden emergency during Christmas time, we needed to rapidly evaluate 1,000 people at risk, so we decided to use conventional serology to detect specific IgM and IgG antibodies via ELISA as well as indirect haemagglutination, which produced positive test results for 9.1%, 11.9% and 9.9% of the individuals tested, respectively. In other more restricted patient groups, polymerase chain reaction (PCR) provided more sensitive results (80.4%) than blood cultures (16.2%) and animal inoculations (11.6%). Although the classical diagnosis of acute ChD is mainly based on parasitological findings, highly sensitive and specific serological techniques can provide rapid results during large and severe outbreaks, as described herein. The use of these serological techniques allows prompt treatment of all individuals suspected of being infected, resulting in reduced rates of morbidity and mortality.

Trypanosoma cruzi I genotypes in different geographical regions and transmission cycles based on a microsatellite motif of the intergenic spacer of spliced-leader genes.

The intergenic region of spliced-leader (SL-IR) genes from 105 Trypanosoma cruzi I (Tc I) infected biological samples, culture isolates and stocks from 11 endemic countries, from Argentina to the USA were characterised, allowing identification of 76 genotypes with 54 polymorphic sites from 123 aligned sequences. On the basis of the microsatellite motif proposed by Herrera et al. (2007) to define four haplotypes in Colombia, we could classify these genotypes into four distinct Tc I SL-IR groups, three corresponding to the former haplotypes Ia (11 genotypes), Ib (11 genotypes) and Id (35 genotypes); and one novel group, Ie (19 genotypes). Genotypes harbouring the Tc Ic motif were not detected in our study. Tc Ia was associated with domestic cycles in southern and northern South America and sylvatic cycles in Central and North America. Tc Ib was found in all transmission cycles from Colombia. Tc Id was identified in all transmission cycles from Argentina and Colombia, including Chagas cardiomyopathy patients, sylvatic Brazilian samples and human cases from French Guiana, Panama and Venezuela. Tc Ie gathered five samples from domestic Triatoma infestans from northern Argentina, nine samples from wild Mepraia spinolai and Mepraia gajardoi and two chagasic patients from Chile and one from a Bolivian patient with chagasic reactivation. Mixed infections by Tc Ia+Tc Id, Tc Ia+Tc Ie and Tc Id+Tc Ie were detected in vector faeces and isolates from human and vector samples. In addition, Tc Ia and Tc Id were identified in different tissues from a heart transplanted Chagas cardiomyopathy patient with reactivation, denoting histotropism. Trypanosoma cruzi I SL-IR genotypes from parasites infecting Triatoma gerstaeckeri and Didelphis virginiana from USA, T. infestans from Paraguay, Rhodnius nasutus and Rhodnius neglectus from Brazil and M. spinolai and M. gajardoi from Chile are to our knowledge described for the first time.