Mitogenome and Nuclear-encoded Fungicide-target Genes of Thecaphora frezii - Causal Agent of Peanut Smut

Background: Thecaphora frezii Carranza and Lindquist causes smut disease in peanut (Arachis hypogaea L.) resulting in up to 35% yield losses. Fungicides have shown ineffective in controlling the disease; whereas research on the molecular basis of that fungicide resistance has been hindered because of the lack of genetic information about T. frezii. The goal of this work was to provide molecular information about fungicide-target loci in T. frezii, including its mitochondrial genome (mitogenome) and critical nuclear-encoded genes. Results: Here we report the complete annotated mitogenome of T. frezii, a 123,773 bp molecule containing the standard 14 genes that form part of mitochondrial complexes I, III, IV and V, 22 transfer RNAs, small and large subunits of ribosomal RNA, DNA polymerase, ribonuclease P, GII-reverse transcriptase/maturase, nine hypothetical open-reading frames and homing endonucleases (LAGLIDADG, GIY-YIG, HEG). In addition, we report the full-length cDNA sequence of T. frezii cytochrome b (cob) and cytochrome oxidase 1 (cox1) genes; as well as partial sequences of T. frezii succinate dehydrogenase (sdhb), ergosterol biosynthesis (Erg4), cytochrome P450 (cyp51), and beta tubulin (β-tubulin) genes, which are respective targets of strobilurins, quinone oxidation inhibitors, triazoles and beta-tubulin inhibitor fungicides commonly used in the peanut crop. Translation of cob and sdhb genes in this particular T. frezii isolate suggests potential resistance to strobilurin and carboxamide fungicides. Conclusion: The mitogenome and nuclear-encoded gene sequences presented here provide the molecular tools to research T. frezii fungicide-target loci

CNVs in the 22q11.2 chromosomal region should be an early suspect in infants with congenital cardiac disease

Q2Background: Congenital heart disease (CHD) is the most common congenital malformation, it is frequently found as an isolated defect, and the etiology is not completely understood. Although most of the cases have multifactorial causes, they can also be secondary to chromosomal abnormalities, monogenic diseases, microduplications or microdeletions, among others. Copy number variations (CNVs) at 22q11.2 are associated with a variety of symptoms including CHD, thymic aplasia, and developmental and behavioral manifestations. We tested CNVs in the 22q11.2 chromosomal region by MLPA in a cohort of Colombian patients with isolated CHD to establish the frequency of these CNVs in the cohort. Methods: CNVs analysis of 22q11.2 by MLPA were performed in 32 patients with apparently isolate CHD during the neonatal period. Participants were enrolled from different hospitals in Bogotá, and they underwent a clinical assessment by a cardiologist and a clinical geneticist. Results: CNVs in the 22q11.2 chromosomal region were found in 7 patients (21.9%). The typical deletion was found in 6 patients (18.75%) and atypical 1.5 Mb duplication was found in 1 patient (3.1%). Conclusions: CNVs in 22q11.2 is a common finding in patients presenting with isolated congenital cardiac disease, therefore these patients should be tested early despite the absence of other clinical manifestations. MLPA is a very useful molecular method and provides an accurate diagnosis.Revista Internacional - Indexad

ACTIVIDAD ANTIFUNGICA DE QUITOSANO CARBAMATO DE ETILO EN Candida albicans

El incremento de las infecciones producidas en seres humanos y/o animales por hongos y el aumento de la resistencia de estos últimos a algunos fármacos, han conducido a la búsqueda de nuevos productos con actividad antifúngica.Producto de la síntesis química se ha obtenido la sustancia de origen natural, llamada Quitosano Carbamato de etilo (QCE) que ha demostrado acción antifúngica, particularmente a Candida albicans. Los objetivos de este trabajo fueron determinar “in vitro” la Concentración MínimaInhibitoria (CIM), Concentración Letal Media (CLM) y Letal (CL) de QCE para cultivos de C. albicans en CM al 2%. Establecer para esta misma levadura su curva de crecimiento, el tiempo de inicio de acción del QCE y los cambios morfológicos de las células levaduriformes en CM al 2% con adición de QCE (0.89 mg/mL). Además, determinar la acción del QCE (0.89 mg/mL) para las levaduras Criptococcus albus, Saccharomyces cerevisiae, Sporobolomyces roseus y 5 cepas de C. albicans aisladasde casos clínicos en el Hospital Clínico Regional Valdivia (H.C.R.V.).La CIM fue 0.25 mg/mL, la CLM fue 0.45 mg/mL y la CL 0,89 mg/mL. La curva de crecimiento elaborada para los cultivos de C. albicans sin adición de QCE fue la típica de una levadura y, por el contrario, la elaborada para los cultivos adicionados con QCE sólo registró laetapa de latencia. El tiempo de inicio de la acción del QCE para la cepa control de C. albicans comienza a las 12 horas de incubación. Las células de C. albicans obtenidas de cultivos tratados con QCE se mostraron colapsadas, de menor tamaño, no forman tubo germinativo y tampocopseudomicelio. El QCE inhibió el desarrollo de cultivos de C. albus, S. cerevisiae, S. roseus y para tres de cinco cepas ensayadas de C. albicans obtenidas desde casos clínicos

Vertical transmission of a Drosophila endosymbiont via cooption of the yolk transport and internalization machinery

Spiroplasma is a diverse bacterial clade that includes many vertically transmitted insect endosymbionts, including Spiroplasma poulsonii, a natural endosymbiont of Drosophila melanogaster. These bacteria persist in the hemolymph of their adult host and exhibit efficient vertical transmission from mother to offspring. In this study, we analyzed the mechanism that underlies their vertical transmission, and here we provide strong evidence that these bacteria use the yolk uptake machinery to colonize the germ line. We show that Spiroplasma reaches the oocyte by passing through the intercellular space surrounding the ovarian follicle cells and is then endocytosed into oocytes within yolk granules during the vitellogenic stages of oogenesis. Mutations that disrupt yolk uptake by oocytes inhibit vertical Spiroplasma transmission and lead to an accumulation of these bacteria outside the oocyte. Impairment of yolk secretion by the fat body results in Spiroplasma not reaching the oocyte and a severe reduction of vertical transmission. We propose a model in which Spiroplasma first interacts with yolk in the hemolymph to gain access to the oocyte and then uses the yolk receptor, Yolkless, to be endocytosed into the oocyte. Cooption of the yolk uptake machinery is a powerful strategy for endosymbionts to target the germ line and achieve vertical transmission. This mechanism may apply to other endosymbionts and provides a possible explanation for endosymbiont host specificity. IMPORTANCE: Most insect species, including important disease vectors and crop pests, harbor vertically transmitted endosymbiotic bacteria. Studies have shown that many facultative endosymbionts, including Spiroplasma, confer protection against different classes of parasites on their hosts and therefore are attractive tools for the control of vector-borne diseases. The ability to be efficiently transmitted from females to their offspring is the key feature shaping associations between insects and their inherited endosymbionts, but to date, little is known about the mechanisms involved. In oviparous animals, yolk accumulates in developing eggs and serves to meet the nutritional demands of embryonic development. Here we show that Spiroplasma coopts the yolk transport and uptake machinery to colonize the germ line and ensure efficient vertical transmission. The uptake of yolk is a female germ line-specific feature and therefore an attractive target for cooption by endosymbionts that need to maintain high-fidelity maternal transmission

The Role of Lipid Competition for Endosymbiont-Mediated Protection against Parasitoid Wasps in Drosophila

Insects commonly harbor facultative bacterial endosymbionts, such as Wolbachia and Spiroplasma species, that are vertically transmitted from mothers to their offspring. These endosymbiontic bacteria increase their propagation by manipulating host reproduction or by protecting their hosts against natural enemies. While an increasing number of studies have reported endosymbiont-mediated protection, little is known about the mechanisms underlying this protection. Here, we analyze the mechanisms underlying protection from parasitoid wasps in Drosophila melanogaster mediated by its facultative endosymbiont Spiroplasma poulsonii. Our results indicate that S. poulsonii exerts protection against two distantly related wasp species, Leptopilina boulardi and Asobara tabida. S. poulsonii-mediated protection against parasitoid wasps takes place at the pupal stage and is not associated with an increased cellular immune response. In this work, we provide three important observations that support the notion that S. poulsonii bacteria and wasp larvae compete for host lipids and that this competition underlies symbiont-mediated protection. First, lipid quantification shows that both S. poulsonii and parasitoid wasps deplete D. melanogaster hemolymph lipids. Second, the depletion of hemolymphatic lipids using the Lpp RNA interference (Lpp RNAi) construct reduces wasp success in larvae that are not infected with S. poulsonii and blocks S. poulsonii growth. Third, we show that the growth of S. poulsonii bacteria is not affected by the presence of the wasps, indicating that when S. poulsonii is present, larval wasps will develop in a lipid-depleted environment. We propose that competition for host lipids may be relevant to endosymbiont-mediated protection in other systems and could explain the broad spectrum of protection provide

Negative Regulation by Amidase PGRPs Shapes the Drosophila Antibacterial Response and Protects the Fly from Innocuous Infection

Peptidoglycan recognition proteins (PGRPs) are key regulators of insect immune responses. In addition to recognition PGRPs, which activate the Toll and Imd pathways, the Drosophila genome encodes six catalytic PGRPs with the capacity to scavenge peptidoglycan. We have performed a systematic analysis of catalytic PGRP function using deletions, separately and in combination. Our findings support the role of PGRP-LB as a negative regulator of the Imd pathway and brought to light a synergy of PGRP-SCs with PGRP-LB in the systemic response. Flies lacking all six catalytic PGRPs were still viable but exhibited deleterious immune responses to innocuous gut infections. Together with recent studies on mammalian PGRPs, our study uncovers a conserved role for PGRPs in gut homeostasis. Analysis of the immune phenotype of flies lacking all catalytic PGRPs and the Imd regulator Pirk reveals that the Imd-mediated immune response is highly constrained by the existence of multiple negative feedbacks

Isolation, Characterization, and Biotechnological Potential of Native Microalgae From the Peruvian Amazon

The objective of this chapter is to provide scientific information on basic aspects to be taken into account to achieve the successful isolation, biochemical, and molecular characterization and then to evaluate the biotechnological potential of native microalgae of the Peruvian Amazon. Recent investigations reported by our research team has demonstrated that the isolated native microalgae from the Peruvian Amazon have a great potential for the biotechnological production of biodiesel and nutraceuticals. This biotechnological potential was identified thanks to the application of various protocols that were standardized by the authors over the last 5 years. In conclusion, the native microalgae of the Peruvian Amazon have biotechnological potential and are therefore promising for the production of both biodiesel and nutraceuticals. Various species of microalgae were identified, isolated, cultured, and characterized using biochemical, nutraceutical, and molecular techniques, the isolation stage being the starting point to achieve various biotechnological applications. Ankistrodesmus sp. is one of the microalgae with potential for the production of biodiesel and microalgae such as Haematococcus pluvialis, Scenedesmus sp., and Chlorella sp., among others demonstrated a high potential for nutraceutical production. The stress conditions to which microalgae are subjected are being a determining factor for the production of biodiesel and nutraceuticals

Loss and gain of gut bacterial phylotype symbionts in afrotropical stingless bee species (Apidae: Meliponinae)

Stingless bees (Apidae: Meliponini) are the most diverse group of corbiculate bees and are important managed and wild pollinators distributed in the tropical and subtropical regions of the globe. However, little is known about their associated beneficial microbes that play major roles in host nutrition, detoxification, growth, activation of immune responses, and protection against pathogens in their sister groups, honeybees and bumble bees. Here, we provide an initial characterization of the gut bacterial microbiota of eight stingless bee species from sub-Saharan Africa using 16S rRNA amplicon sequencing. Our findings revealed that Firmicutes, Actinobacteria, and Proteobacteria were the dominant and conserved phyla across the eight stingless bee species. Additionally, we found significant geographical and host intra-species-specific bacterial diversity. Notably, African strains showed significant phylogenetic clustering when compared with strains from other continents, and each stingless bee species has its own microbial composition with its own dominant bacterial genus. Our results suggest host selective mechanisms maintain distinct gut communities among sympatric species and thus constitute an important resource for future studies on bee health management and host-microbe co-evolution and adaptation.Figure S1: Alpha rarefaction curve. Figure S2: Gut bacterial genera associated with each of the eight stingless bee species in Kenya. Figure S3: Bacterial alpha diversity did not correlate with the stingless bee size. Table S1: 16S rRNA sequencing data analysis (Excel file).Supplementary material: Figure S1: Alpha rarefaction curve. Figure S2: Gut bacterial genera associated with each of the eight stingless bee species in Kenya. Figure S3: Bacterial alpha diversity did not correlate with the stingless bee size. Table S1: 16S rRNA sequencing data analysis (Excel file).The Swiss National Science Foundation Postdoc Mobility, the German Academic Exchange Service (DAAD), Norwegian Agency for Development Cooperation, the Section for Research, Innovation, and Higher Education and by icipe core donors.https://www.mdpi.com/journal/microorganismsam2022BiochemistryForestry and Agricultural Biotechnology Institute (FABI)GeneticsMicrobiology and Plant Patholog

Effect of different housing systems (single and group penning) on the health and welfare of commercial female rabbits

[EN] In recent decades, concern about rabbit welfare and sustainability has increased. The housing system is a very important factor for animal welfare. However, information about how different available housing types for female rabbits affect their health status is scarce, but this is an important factor for their welfare. Hence, the objective of this study was to evaluate the health status of female rabbits in five common housing systems: three different single-housing systems with distinct available surfaces and heights; a single-housing system with a platform; a collective system. Female rabbits in the collective and platform cages had greater cortisol concentrations in hair than those in the single-housing system with no platform. Haptoglobin concentrations and kit mortality rates during lactation were greater for the collective-cage female rabbits. The collective group had more culled females and more lesions than in the other groups. The main reasons for culling in all the groups were reproduction problems and presence of abscesses, and the collective group of females was the most affected. In conclusion, it appears that keeping females together in collective systems negatively affects their health status and welfare, while single-housing systems imply lower kit mortality rates during lactation and cortisol concentrations, and fewer lesions in female rabbits.This study was supported by the INIA-INTERCUN Project (CUN2014-00001-00-00) and by Universidad CEU-Cardenal Herrera (INDI16/07, INDI17/07, INDI18/08). Grants for Sara Perez Fuentes, Asuncion Munoz Silvestre and Elena Moreno Grua from Universidad CEU-Cardenal Herrera, and also for Sara Perez Fuentes from Generalitat Valenciana and FSE (ACIF/2016/085), and for Elena Moreno Grua from Ministerio de Educacion, Cultura y Deporte (FPU17/02708) are also gratefully acknowledged.Pérez-Fuentes, S.; Muñoz-Silvestre, A.; Moreno Grua, E.; Martinez-Paredes, E.; Viana, D.; Selva, L.; Villagrá, A.... (2020). Effect of different housing systems (single and group penning) on the health and welfare of commercial female rabbits. Animal. 14(6):1270-1277. https://doi.org/10.1017/S1751731119003379S12701277146Archetti I, Tittarelli C, Cerioli M, Brivio R, Grilli G and Lavazza A 2008. Serum chemistry and hematology values in commercial rabbits: preliminary data from industrial farms in northern Italy. In Proceedings of the 9th World Rabbit Congress, 10–13 June 2008, Verone, Italy, pp. 1147–1152.Argente, M.-J., García, M. de la L., Birlanga, V., & Muelas, R. (2014). Relationship between cortisol and acute phase protein concentrations in female rabbits. The Veterinary Journal, 202(1), 172-175. doi:10.1016/j.tvjl.2014.07.020Cervera C, Martínez-Paredes E, Machado L and Villagrá A 2017. Producción de conejas en sistemas de alojamiento individual o colectivo en semigrupo. In Proceedings of the XLII Symposium de Cunicultura de ASESCU, 11–12 May 2017, Murcia, Spain, pp. 107–110.Dal Bosco, A., Mugnai, C., Martino, M., Szendrő, Z., Mattioli, S., Cambiotti, V., … Castellini, C. (2019). Housing Rabbit Does in a Combi System with Removable Walls: Effect on Behaviour and Reproductive Performance. Animals, 9(8), 528. doi:10.3390/ani9080528Dalle Zotte, A., Princz, Z., Matics, Z., Gerencsér, Z., Metzger, S., & Szendrő, Z. (2009). Rabbit preference for cages and pens with or without mirrors. Applied Animal Behaviour Science, 116(2-4), 273-278. doi:10.1016/j.applanim.2008.08.011Machado L, Cervera C, Martínez-Paredes E, Paragliola F and Cano C 2016. Comportamiento y manejo de conejas en sistemas de cría colectiva. In Proceedings of the XLI Symposium de Cunicultura de ASESCU, 12–13 May 2016, Hondarribia, Spain, pp. 134–137.Maertens L and Buijs S 2016. Impact of housing system (cage vs. part-time housing) and floor type on rabbit doe welfare. In Proceedings of the 11th World Rabbit Congress, 15–18 June 2016, Qingdao, China, pp. 707–710.Marai, I. F. ., Habeeb, A. A. ., & Gad, A. . (2002). Rabbits’ productive, reproductive and physiological performance traits as affected by heat stress: a review. Livestock Production Science, 78(2), 71-90. doi:10.1016/s0301-6226(02)00091-xMarcato PS and Rosmini R 1986. Mammary glands. In Pathology of the rabbit and hare (eds. PS Marcato and R Rosmini ), pp. 179–186. Società Editrice Esculapio, Bologna, Italy.Masthoff, T., & Hoy, S. (2019). Investigations on the Influence of Floor Design on Dirtiness and Foot Pad Lesions in Growing Rabbits. Animals, 9(6), 354. doi:10.3390/ani9060354Mugnai, C., Dal Bosco, A., & Castellini, C. (2009). Effect of different rearing systems and pre-kindling handling on behaviour and performance of rabbit does. Applied Animal Behaviour Science, 118(1-2), 91-100. doi:10.1016/j.applanim.2009.02.007Petersen, H. H., Nielsen, J. P., & Heegaard, P. M. H. (2004). Application of acute phase protein measurements in veterinary clinical chemistry. Veterinary Research, 35(2), 163-187. doi:10.1051/vetres:2004002Rödel, H. G., Starkloff, A., Bautista, A., Friedrich, A.-C., & Von Holst, D. (2008). Infanticide and Maternal Offspring Defence in European Rabbits under Natural Breeding Conditions. Ethology, 114(1), 22-31. doi:10.1111/j.1439-0310.2007.01447.xRödel, H. G., Starkloff, A., Seltmann, M. W., Prager, G., & von Holst, D. (2009). Causes and predictors of nest mortality in a European rabbit population. Mammalian Biology, 74(3), 198-209. doi:10.1016/j.mambio.2008.04.003Rommers, J. M., Boiti, C., De Jong, I., & Brecchia, G. (2006). Performance and behaviour of rabbit does in a group-housing system with natural mating or artificial insemination. Reproduction Nutrition Development, 46(6), 677-687. doi:10.1051/rnd:2006038Rommers, J., & De Greef, K. H. (2018). Are combi parks just as useful as regular parks for fatteners for part-time group housing of rabbit does? World Rabbit Science, 26(4), 299. doi:10.4995/wrs.2018.9587Rosell J and De La Fuente L 2008. Health and body condition of rabbit does on commercial farms. In Proceedings of the 9th World Rabbit Congress, 10–13 June 2008, Verone, Italy, pp. 1065–1069.Rosell, J., & de la Fuente, L. (2013). Assessing Ulcerative Pododermatitis of Breeding Rabbits. Animals, 3(2), 318-326. doi:10.3390/ani3020318Sánchez, J. P., de la Fuente, L. F., & Rosell, J. M. (2012). Health and body condition of lactating females on rabbit farms1. Journal of Animal Science, 90(7), 2353-2361. doi:10.2527/jas.2011-4065Segura, P., Martinez, J., Peris, B., Selva, L., Viana, D., Penades, J. R., & Corpa, J. M. (2007). Staphylococcal infections in rabbit does on two industrial farms. Veterinary Record, 160(25), 869-872. doi:10.1136/vr.160.25.869Siemsen DW, Malachowa N, Schepetkin IA, Whitney AR, Kirpotina LN, Lei B, DeLeo FR and Quinn MT 2014. Neutrophil isolation from nonhuman species. In Neutrophil methods and protocols (eds. MT Quinn and FR De Leo ), pp. 19–37. Humana Press, Totowa, NJ, USASzendrő, Z., & McNitt, J. I. (2012). Housing of rabbit does: Group and individual systems: A review. Livestock Science, 150(1-3), 1-10. doi:10.1016/j.livsci.2012.09.017Szendrő, Z., Mikó, A., Odermatt, M., Gerencsér, Z., Radnai, I., Dezséry, B., … Matics, Z. (2013). Comparison of performance and welfare of single-caged and group-housed rabbit does. 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Rearing management of rabbit males selected by high growth rate: the effect of diet and season on semen characteristics

[EN] A total of sixty-six young males were used to evaluate the effect of low (L), medium (M) and high (H) concentrations of dietary digestible energy received during the rearing seasons (autumn and spring) on the performance and main semen characteristics of males for artificial insemination selected by a high growth rate. Males reared during the spring season presented a significantly higher weight at weaning than those reared during the autumn season (P < 0.001), and these differences were maintained until the end of the trial. The requirements of the males were easily covered as a general rule. In the autumn group, the males were unable to intake the digestible protein recommended only during their 3rd month of life, especially with low concentrate diets (P < 0.05). H males showed higher semen concentration and production during the autumn season, while L males showed a higher semen concentration and production than M males during the spring season, the H group showed intermediate values (P < 0.001). Males reared during the spring season showed significantly higher values of sperm concentration (P < 0.01) and production (P < 0.01). H males presented a lower percentage of spermatozoa with cytoplasmic droplets than the L group (P < 0.05) and the lowest values for sperm abnormalities during the autumn season, while the L group presented higher values for percentage abnormalities, especially during the last month controlled (P < 0.05). As a general rule, the main motility parameters controlled were not affected by the rearing diet received nor the season. These results seem to indicate that the management of rabbit males during the growing and rearing periods seem to significantly affect their subsequent performance and semen production.The present work was supported by a grant from CICYT: AGF98-0470-C02-01Pascual Amorós, JJ.; García, C.; Martinez-Paredes, E.; Moce Cervera, ET.; Vicente Antón, JS. (2004). Rearing management of rabbit males selected by high growth rate: the effect of diet and season on semen characteristics. Reproduction Nutrition Development. 44(1):49-63. https://doi.org/10.1051/rnd:2004016S496344