The vicuna (Vicugna vicugna) as a natural host of Dictyocaulus filaria in Peru.

Lungworm infection, or verminous pneumonia, is a parasitic disease that causes serious problems in small and large ruminants. Despite the fact that nematodes of the genus Dictyocaulus in cattle and sheep are the main cause of this disease, there are few studies on the natural infections of South American camelids. For this reason, this study aims to report the natural infection by Dictyocaulus filaria in vicunas (Vicugna vicugna) for the first time. During a shearing season (chaku) in Cuzco, Peru, two accidentally killed adult vicunas were submitted to the IVITA-Marangani research center in Cuzco for their respective necropsies. The tracheas of both vicunas had numerous nematodes, as seen during the necropsy. The nematodes were collected in 70% ethanol and were morphologically identified as D. filaria. Likewise, the DNA of six nematodes was extracted, and the ITS2 region and the 28S rRNA gene were amplified and sequenced. The nucleotide sequences of both genetic markers were up to 100% identical with previously reported D. filaria DNA sequences found in the goat yearlings from Turkey, sheep from Iran, Turkey, and India, and the argali from Uzbekistan, which confirmed the morphological diagnosis. This finding represents the first molecular confirmation of a natural D. filaria infection in a South American camelid. It will be necessary to carry out future studies to know the current situation of verminous pneumonia in domestic and wild South American camelids and to know the negative effects of the disease on them.

Identification of wild rodents as intermediate hosts for Hydatigera taeniaeformis in Peru.

Hydatigera taeniaeformis is a cestode that uses felines and rodents as definitive and intermediate hosts, respectively. Its larval stage, or metacestode, infects a wide variety of rodent species and develops in the liver parenchyma into a cyst. The aim of this study was to evaluate the occurrence of H. taeniaeformis metacestode in various species of wild rodents from Peru. For this, the livers of 356 rodents were macroscopically examined for any parasitic form compatible with metacestodes. Metacestodes were identified by measuring characteristic morphological parameters, and the diagnosis was confirmed by molecular analysis of a fragment of the cytochrome c oxidase subunit 1 gene (cox1). Five rodents: two small-eared pygmy rice rats (Oligoryzomys microtis), two white-naped squirrels (Simosciurus nebouxii), and one pygmy rice rat (Oligoryzomys sp.) were infected with H. taeniaeformis metacestodes. The cox1 sequences from our metacestodes showed up to 100% identity with previous H. taeniaeformis sequences from the GenBank. These results demonstrated the occurrence of H. taeniaeformis in new intermediate hosts, as well as the first molecular contribution for H. taeniaeformis from Peru.

Observation and Mitigation of Leachables from Non-Product Contact Materials in Electromechanical Delivery Devices for Biotechnology Products.

Battery-powered drug delivery devices are widely used as primary containers for storing and delivering therapeutic protein products to improve patient compliance and quality of life. Compared to conventional delivery approaches such as pre-filled syringes, battery-powered devices are more complex in design requiring new materials/components for proper functionality, which could cause potential product safety and quality concerns from the extractable and leachables (E&L) of the new materials/components. In this study, E&L assessments were performed on a battery-powered delivery device during the development and qualification of the device, where novel compound 2­hydroxy-2-methylpropiophenone (HMPP) and related compounds were observed in both E&L. The source of the HMPP and related compounds was identified to be the nonproduct contact device batteries, in which HMPP photo-initiator was used as a curing agent in the battery sealant to prevent leakage of the battery electrolytes. Toxicology assessment was performed, which showed the levels of HMPP observed in the device lots were acceptable relative to the permitted daily exposure. A drug product HMPP spike study was also performed, where no product impact was observed. Based on these assessments, an action threshold and specification limits could be established as a control strategy, if needed, to mitigate the potential risks associate with the observed leachables. As a full resolution, seven battery candidates from different suppliers were screened and one new battery was successfully qualified for the delivery devices. Overall, the holistic E&L approach was fully successful in the development and qualification of the battery-powered devices for biotherapeutic products delivery ensuring product quality and patient safety. Non-product contact materials are commonly rated as low or no risk and typically considered as out of scope of E&L activities for delivery systems following industry benchmark and regulatory agency guidance. This case study is novel as it brings into attention the materials that might not normally be in consideration during the development process. It is highly recommended to understand materials in the context of intended use on a case-by-case basis and not to generalize to ensure successful development and qualification.

Review: Mixed-Matrix Membranes with CNT for CO2 Separation Processes.

The membranes' role is of supreme importance in the separation of compounds under different phases of matter. The topic addressed here is based on the use of membranes on the gases separation, specifically the advantages of mixed-matrix membranes (MMMs) when using carbon nanotubes as fillers to separate carbon dioxide (CO2) from other carrier gas. MMMs consist of a polymer support with additive fillers to improve their efficiency by increasing both selectivity and permeability. The most promising fillers in the MMM development are nanostructured molecules. Due to the good prospects of carbon nanotubes (CNTs) as MMM fillers, this article aims to concentrate the advances and developments of CNT-MMM to separate gases, such as CO2. The influence of functionalized CNT or mixtures of CNT with additional materials such as zeolites, hydrogel and, graphene sheets on membranes performance is highlighted in the present work.

Prevalence and molecular characterization of Sarcoptes scabiei from vicuñas (Vicugna vicugna) from Southern Peruvian Andes.

Sarcoptic mange is a disease caused by an infectious parasite in the vicuñas (Vicugna vicugna) from South America. Although molecular studies have provided much information about the epidemiology of this disease, this information is still unknown in vicuñas. This study determined the prevalence and molecular characterization of Sarcoptes scabiei from vicuñas from Southern Peruvian Andes. During the 2018 shearing season, 181 vicuñas were clinically evaluated for lesions compatible with mange. Sarcoptes scabiei was detected in 35 (19.3%) vicuñas, and 50 mites from 25 vicuñas were selected for molecular analyses of the mitochondrial (cox1) and nuclear (ITS2) genetic markers. Molecular analyses of the cox1 and ITS2 sequences showed an identity of 94­99% and 99.8­100% with previous S. scabiei sequences registered in the GenBank, respectively. Sequence polymorphisms were more evident in the ITS2 than in the cox1, but only the cox1 had an association with the host. Phylogenetic analysis of S. scabiei cox1 sequences from vicuñas showed a cluster with S. scabiei cox1 sequences from canids, suggesting that the origin of S. scabiei from vicuña is associated with canid mites. This research is the first molecular analysis of S. scabiei from vicuñas. Future molecular studies will be necessary to determine the species variety, geographic segregation and host­parasite adaptation for this vicuña's mite.

Modifications of extracellular matrix features in the left and right uterine horns during the embryo pre-implantation period in Vicugna pacos.

More than 98% of the pregnancies in South American camelids is carried out in the left uterine horn (LUH). Hence, embryos originated from right-ovary ovulations have to migrate to the contralateral or left uterine horn (LUH) to implant and survive. A reason for this unique pattern of embryo implantation has not been elucidated yet. In general, embryo implantation involves an extensive extracellular matrix (ECM) remodeling within the endometrium, in which collagen and matrix metalloproteinases (MMPs) play an essential role. Deregulation of collagen and MMPs has been related to embryo implantation failure, miscarriage, and infertility. Therefore, we hypothesized that ECM components in camelids could be involved in differential embryo implantation and consequently the high incidence of left horn gestations. The aim of this study was to describe and compare changes in ECM components in the left and right uterine horn of non-pregnant and 15 days pregnant alpacas. To test this hypothesis, the collagen content was evaluated by specific staining with Picrosirius Red and using ImageJ 1.42q software. Subsequently, gene expression of the following components of the MMP pathway was determined: MMP-2, -3, -7, -9, and -14, MMP substrates (COL1A2 and COL3A1), MMP inhibitors (TIMP1 and TIMP2), LGMN, an MMP activator, and EMMPRIN, an extracellular matrix metalloproteinase inducer. Uterine horns of pregnant alpacas exhibited a marked decrease in collagen content. In contrast, transcript expression of COL1A2 and COL3A1 was higher in the LUH of pregnant alpacas. Gene expression of MMP-3, -7, -9, -14, LGMN, and EMMPRIN were also higher in the LUH of pregnant animals, whereas MMP-2 gene expression was higher in the LUH of both pregnant and non-pregnant alpacas. Expression of TIMP1 and TIMP2 increased during pregnancy, with higher values in the LUH. In conclusion, expression of ECM components displayed a specific pattern depending on the uterine side and the physiological status (pregnant vs non-pregnant) of the animal. The increased expression of ECM transcripts in the left uterine horn during early pregnancy in alpacas suggests the involvement of these molecules in a highly regulated process leading to the implantation process.

Infección natural por Fasciola hepatica en cérvidos del Perú / Natural infection of Fasciola hepatica in cervids from Peru

En el presente trabajo se registra la infección natural por Fasciola hepatica en un venado de cola blanca (Odocoileus virginianus) y en una taruca (Hippocamelus antisensis), ambos procedentes del departamento de Cusco. Los animales fueron remitidos al Instituto Veterinario (IVITA-Maranganí, FMV, UNMSM) por las autoridades del Servicio Nacional de Flora y Fauna (SERFOR, Sede Cusco). Durante la necropsia de los animales se colectaron seis trematodos de los conductos biliares, los cuales fueron preservados en etanol al 70%. Las observaciones morfológicas indicaron que se trataban de F. hepatica. Esto fue confirmado analizando el ADN mitocondrial de los parásitos amplificando parcialmente los genes citocromo c oxidasa subunidad 1 (cox1) y el NADH deshidrogenasa subunidad 1 (nad1). El análisis de estos genes tuvo una identidad mayor al 99% comparado con registros del banco de genes (GenBank). El presente estudio demuestra la presencia de F. hepatica en estos cérvidos, agregando así dos nuevos hospederos definitivos para el parásito.

Natural infection by Fasciola hepatica is recorded in a white-tailed deer (Odocoileus virginianus) and a taruca (Hippocamelus antisensis), both from the department of Cusco. Animals were remitted to the Veterinary Institute (IVITA-Maranganí, FMV, UNMSM) by the authorities of the National Service of Flora and Fauna (SERFOR, Cusco Headquarters). Six trematodes were collected from the bile ducts during the necropsy of the animals, and they were preserved in 70% ethanol. Morphological analysis indicated that they correspond to F. hepatica. This was confirmed by analyzing of the mitochondrial DNA of the parasites by partially amplifying the cytochrome c oxidase subunit 1 (cox1) and the NADH dehydrogenase subunit 1 (nad1) genes. Analysis of these genes had an identity greater than 99% compared to genes from GenBank. The present study demonstrates the occurrence of F. hepatica in these cervids, thus adding two new definitive hosts for the parasite.

Changes in mucins and matrix metalloproteases in the endometrium of early pregnant alpacas (Vicugna pacos).

South American Camelids (SAC) have unique reproductive features, one of which is that 98% of the pregnancies develop in the left uterine horn. Furthermore, early pregnancy is an uncharacterized process in these species, especially in regard to the ultrastructural, biochemical and genetic changes that the uterine epithelial surface undergoes to allow embryo implantation. The present study describes the uterine horn luminal surface and the characteristics of the mucinous glycocalyx in non-pregnant and early pregnant (15 days) female alpacas. In addition, the relative abundance of Mucin 1 and 16 genes (MUC1 and MUC16) was determined, as well as the relative mRNA abundance of matrix metalloproteinases (MMPs) that could be involved in MUC shedding during early pregnancy. Noticeable changes were detected in the uterine luminal epithelium and glycocalyx of pregnant alpacas in comparison to non-pregnant ones, as well as presence of MUCs and MMPs in the endometrial environment. The decrease in glycocalyx staining and in the relative abundance of MUC 1 and MUC 16 transcripts in pregnant females would allow embryo attachment to the luminal epithelium and its subsequent implantation, as has been described in other mammals. These results suggest a crucial role of MUC1 and MUC16 and a possible role of MMPs in successful embryo implantation and survival in alpacas.

First finding of nymphal stages of Linguatula serrata in a South American camelid, a vicuña from Peru.

Linguatula serrata, a pentastomid, was found parasitizing the lungs of a vicuña (Vicugna vicugna) from Cuzco, Peru. A total of 13 larvae were found encysted in the parenchymal tissue of the lungs. All larvae were identified as nymphal stages of L. serrata by morphological methods Diagnosis was confirmed by molecular analysis amplifying the cytochrome c oxidase 1 gene of three nymphs. Nucleotide sequences from the isolates were compared to previous sequences from GenBank, and it showed high similarity between them (>99%). This finding constitutes the first detection of L. serrata in a South American camelid.

Molecular and morphological evidence of Taenia omissa in pumas (Puma concolor) in the Peruvian Highlands.

A total of 41 cestodes were collected during necropsy examination on 2 pumas (Puma concolor) that were found in 2 communities in Canchis province, Cuzco region, Peru, at 4500 meters above sea level (Peruvian Andes). The cestodes were evaluated morphologically and molecularly. A fragment of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) was used as a genetic marker. All the cestodes were identified as Taenia omissa. In the present report, we give a brief description by molecular and morphological diagnosis of the cestodes and compare nucleotide sequences with previous isolates from GenBank. Upon comparison, the sequences showed a difference in the cox1 gene of 5.1 to 5.3% with other teniids sequences. This finding constitutes the first report of T. omissa in Peru and expands the geographic distribution of this parasite.

Sobre algunos helmintos parásitos de la taruca, Hippocamelus antisensis (Mammalia: Artiodactyla) / On some helminth parasites of the taruca, Hippocamelus antisensis (Mammalia: Artiodactyla)

En el presente estudio se da a conocer tres especies de helmintos parásitos colectados de una taruca (Hippocamelus antisensis) en el Perú. Una taruca macho adulto fue remitido al Instituto Veterinario de Investigaciones Tropicales y de Altura (IVITA), Sede Maranganí, por el Servicio Nacional Forestal y de Fauna Silvestre del Perú (SERFOR), para su respectiva necropsia. Se colectaron 2 nematodos del sistema digestivo, así como un quiste localizado en el omento. Para el diagnóstico parasitológico se evaluaron las características morfo-métricas de los especímenes colectados. Los nematodos fueron diagnosticados como Trichostrongylus axei y Mazamastrongylus sp.; por otro lado, el quiste fue diagnosticado como un metacestodo de Taenia hydatigena. El hallazgo de T. axei representa el primer registro para la taruca. En la presente nota se realiza también una breve descripción morfológica de los helmintos mencionados.

This study presents three species of helminth parasites collected from a taruca (Hippocamelus antisensis) in Peru. Three helminth parasites from the taruca (Hippocamelus antisensis) are described in this report. An adult male taruca was remitted to the Veterinary Institute of Tropical and Highland Research (IVITA), Sede Maranganí, for the National Forest and Wildlife Service of Peru (SERFOR) for their respective necropsy. Two nematodes were collected from the digestive system, and 1 cyst was collected from the omentum. Morphometric characteristics of the specimens were evaluated for the parasitological diagnosis. Nematodes were diagnosed as Trichostrongylus axei and Mazamastrongylus sp.; on the other hand, the cyst was diagnosed as a metacestode of Taenia hydatigena. T. axei finding represents the first record for the taruca. A brief morphological description of each helminth and discussion about their hosts and distribution is done.

Molecular and morphological evidence of Taenia omissa in pumas (Puma concolor) in the Peruvian Highlands / Evidência molecular e morfológica de Taenia omissa em onça-pardas (Puma concolor) dos Andes Peruanos

Abstract A total of 41 cestodes were collected during necropsy examination on 2 pumas (Puma concolor) that were found in 2 communities in Canchis province, Cuzco region, Peru, at 4500 meters above sea level (Peruvian Andes). The cestodes were evaluated morphologically and molecularly. A fragment of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) was used as a genetic marker. All the cestodes were identified as Taenia omissa. In the present report, we give a brief description by molecular and morphological diagnosis of the cestodes and compare nucleotide sequences with previous isolates from GenBank. Upon comparison, the sequences showed a difference in the cox1 gene of 5.1 to 5.3% with other teniids sequences. This finding constitutes the first report of T. omissa in Peru and expands the geographic distribution of this parasite.

Resumo Um total de quarenta e um cestóides foram coletados durante a necropsia de duas onça-pardas (Puma concolor) encontradas em duas comunidades na província de Canchis, em Cuzco, a 4500 metros acima do nível do mar, nos Andes peruanos. Os cestóides foram avaliados morfologicamente e molecularmente. Um fragmento do gene citocromo C oxidase subunidade 1 (cox1) foi utilizado como marcador genético. Todos os cestóides foram identificados como Taenia omissa. No presente relato, dá-se uma breve descrição dos cestóides e compara-se sequências de nucleotídeos com isolados anteriores presentes no GenBank. Após a comparação, as sequências mostraram uma diferença de 5,1-5,3% entre o gene cox1 e outras sequências de tênias. Esse achado constitui o primeiro relato de T. omissa no Peru e amplia a informação sobre a distribuição geográfica deste parasita.

The taruca (Hippocamelus antisensis) and the red brocket deer (Mazama americana) as intermediate hosts of Taenia hydatigena in Peru, morphological and molecular evidence.

In the present report metacestodes were collected from the mesentery of a taruca (Hippocamelus antisensis) and from the omentum of a red brocket deer (Mazama americana) in Peru. Various metacestodes parameters, including rostellar hook characteristics, were measured. Molecular analysis was performed to amplify the mitochondrial cytochrome c oxidase subunit 1 gene from metacestode isolates. Metacestodes were identified as T. hydatigena by morphology and molecular methods. This constitutes the first molecular detection of T. hydatigena metacestodes in the taruca and the red brocket deer and demonstrates that these animal species are natural intermediate hosts for this parasite.