Lumbriculus variegatus (Müller, 1774) (Oligochaeta, Lumbriculidae) in Campos do Jordão State Park, São Paulo, Brazil / Lumbriculus variegatus (Müller, 1774) (Oligochaeta, Lumbriculidae) no Parque Estadual de Campos do Jordão, São Paulo, Brasil

Abstract The aim of this paper was recording the occurrence of the species Lumbriculus variegatus (Müller, 1774) (Oligochaeta, Lumbriculidae) in lotic systems of the State of São Paulo. Specimens were collected in Sapucaí River, located in Campos do Jordão State Park. The mapping of geographical distribution of this species is of interest to public health since L. variegatus may be an intermediate host of Dioctophyme renale (Goeze, 1782) (Nematoda, Dioctophymatidae), a parasite of recognized zoonotic potential. Distribution data serves as a basis for environmental monitoring and evaluation, being essential to map possible cases of the disease (Dioctophimosis) and provide information to health professionals.

Resumo O objetivo deste trabalho foi registrar a ocorrência de Lumbriculus variegatus (Müller, 1774) (Oligochaeta, Lumbriculidae) em um sistema lótico do Estado de São Paulo. Os espécimes foram coletados no rio Sapucaí, localizado no Parque Estadual de Campos do Jordão. O mapeamento da distribuição geográfica desta espécie é de interesse para saúde pública uma vez que L. variegatus pode ser um hospedeiro intermediário de Dioctophyme renale (Goeze, 1782) (Nematoda, Dioctophymatidae), parasito de reconhecido potencial zoonótico. Dados de distribuição servem de base para monitoramento e avaliação ambiental, sendo essenciais para mapear possíveis casos da doença (Dioctofimose) e fornecer informações para profissionais de saúde.

The impact of cow dung augmentation on soil restoration and bio-accumulation of metals (Lead and Cadmium) in Pheretima posthuma (Annelida: Clitellata) / O impacto do aumento de esterco de vaca na restauração do solo e bioacumulação de metais (chumbo e cádmio) em Pheretima posthuma (Annelida: Clitellata)

Abstract To investigate the role of cow dung in soil reclamation and bio assimilation along with bio accumulation of heavy metals in earthworm (P. posthuma) (N=900) earthworms were used and treatment groups of CD-soil mixture of different proportion of cow dung were designed. Nonlethal doses of lead acetate and cadmium chloride were added in treatment groups. Mature P. posthuma were released in each experimental pot maintaining the favorable conditions. The pH, carbon, nitrogen, phosphorus, exchangeable cations, and heavy metal level of each mixture was evaluated. The results indicated that bio-assimilation of Pb and Cd by P. posthuma were significantly (P ˂ 0.01) higher in different soil-CD treatments compared to control. Highest bio-assimilation of both metals was observed in T1 of both groups (Pb = 563.8 mg/kg and Cd = 42.95 mg/kg). The contents of both metals were significantly (P ˂ 0.05) lowered in casting. The nutrient concentration in the final castings of all soil-CD treatments were also equally transformed from less or insoluble to more soluble and available for plants, except for carbon level which increased with CD proportion. It is concluded that cow dung as organic matter has a positive effect on soil reclamation and bio-assimilation of metals by P. posthuma.

RESUMO Para investigar o papel do esterco de vaca na recuperação do solo e bioassimilação, juntamente com a bioacumulação de metais pesados ​​em minhocas (P. posthuma) (N = 900), minhocas foram usadas e grupos de tratamento de mistura CD-solo de diferentes proporções de esterco de vaca foram projetados. Doses não letais de acetato de chumbo e cloreto de cádmio foram adicionadas aos grupos de tratamento. P. posthuma maduros foram liberados em cada vaso experimental, mantendo as condições favoráveis. Foram avaliados o pH, carbono, nitrogênio, fósforo, cátions trocáveis ​​e nível de metais pesados ​​de cada mistura. Os resultados indicaram que a bioassimilação de Pb e Cd por P. posthuma foi significativamente (P ˂ 0,01) maior em diferentes tratamentos de solo-CD em relação ao controle. A maior bioassimilação de ambos os metais foi observada em T1 de ambos os grupos (Pb = 563,8 mg / kg e Cd = 42,95 mg / kg). O conteúdo de ambos os metais foi significativamente (P ˂ 0,05) reduzido na fundição. A concentração de nutrientes nas fundições finais de todos os tratamentos de solo-CD também foi igualmente transformada de menos ou insolúvel para mais solúvel e disponível para as plantas, exceto o nível de carbono que aumenta com a proporção de CD. Conclui-se que o esterco de vaca como matéria orgânica tem um efeito positivo na recuperação do solo e na bioassimilação de metais por P. posthuma.

Antibacterial and bacteriostatic potential of coelomic fluid and body paste of Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) against ampicillin resistant clinical bacterial isolates / Potencial antibacteriano e bacteriostático do fluido celômico e pasta corporal de Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) contra isolados bacterianos clínicos resistentes à ampicilina

Abstract Pheretima posthuma (Vaillant, 1868), a native earthworm of Pakistan and Southeast Asia, has wide utilization in vermicomposting and bioremediation process. In this study, P. posthuma coelomic fluid (PCF) and body paste (PBP) was evaluated as antibacterial agent against ampicillin (AMP) resistant five Gram positive and four Gram negative clinical isolates. The antibacterial effect of different doses (i.e. 25-100 µg/ml) of PCF and PBP along with AMP and azithromycin (AZM) (negative and positive controls, respectively) were observed through disc diffusion and micro-dilution methods. All nine clinical isolates were noticed as AMP resistant and AZM sensitive. Antibacterial effects of PCF and PBP were dose dependent and zone of inhibitions (ZI) against all clinical isolates were between 23.4 ± 0.92 to 0 ± 00 mm. The sensitivity profile of PCF and PBP against clinical isolates was noticed as 44.44 and 55.56%, respectively. Both PCF and PBP showed bacteriostatic (BTS) action against S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Moreover, the cumulative BTS potential of PCF and PBP against all isolates was 66.67 and 55.56%, respectively. The MICs of PCF and PBP were ranged from 50-200 µg/ml against selected isolates. The bacterial growth curves indicated that PCF and PBP inhibited the growth of all isolates at their specific MIC concentrations. However, PBP has better antibacterial potential compared to PCF against selected isolates. Therefore, it is concluded that both PCF and PBP of P. posthuma possess antibacterial and BTS potential against ampicillin resistant clinical isolates. This organism might be considered as a second choice of antibacterial agents and can further be utilized in pharmaceutical industries for novel drug manufacturing by prospecting bioactive potential agents.

Resumo Pheretima posthuma (Vaillant, 1868), uma minhoca nativa do Paquistão e sudeste da Ásia, tem ampla utilização em processos de vermicompostagem e biorremediação. Neste estudo, o fluido celômico de P. posthuma (PCF) e a pasta corporal (PBP) foram avaliados como agente antibacteriano contra cinco isolados clínicos Gram-positivos e quatro Gram-negativos resistentes à ampicilina (AMP). O efeito antibacteriano de diferentes doses (ou seja, 25-100 µg / ml) de PCF e PBP juntamente com AMP e azitromicina (AZM) (controles negativo e positivo, respectivamente) foi observado por meio de métodos de difusão em disco e microdiluição. Todos os nove isolados clínicos foram notados como resistentes a AMP e sensíveis a AZM. Os efeitos antibacterianos de PCF e PBP foram dependentes da dose e a zona de inibição (ZI) contra todos os isolados clínicos foi entre 23,4 ± 0,92 a 0 ± 00 mm. O perfil de sensibilidade do PCF e PBP contra isolados clínicos foi observado como 44,44% e 55,56%, respectivamente. Tanto o PCF quanto o PBP mostraram ação bacteriostática (BTS) contra S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Além disso, o potencial BTS cumulativo de PCF e PBP contra todos os isolados foi de 66,67% e 55,56%, respectivamente. Os MICs de PCF e PBP variaram de 50-200 µg / ml contra isolados selecionados. As curvas de crescimento bacteriano indicaram que o PCF e o PBP inibiram o crescimento de todos os isolados em suas concentrações específicas de MIC. No entanto, PBP tem melhor potencial antibacteriano em comparação com PCF contra isolados selecionados. Portanto, conclui-se que tanto o PCF quanto o PBP de P. posthuma possuem potencial antibacteriano e BTS contra isolados clínicos resistentes à ampicilina. Esse organismo pode ser considerado como uma segunda escolha de agentes antibacterianos e pode ainda ser utilizado nas indústrias farmacêuticas para a fabricação de novos medicamentos por meio da prospecção de agentes com potencial bioativo.

Histopathological and immunohistochemical study of antiosteoporotic efficacy of the earthworm Allolobophora caliginosa extract in orchiectomized rats / Estudio histopatológico e inmunohistoquímico de la eficacia antiosteoporótica del extracto de lombriz de tierra Allolobophora caliginosa en ratas orquiectomizadas

SUMMARY: Osteoporosis is a bone condition marked by a loss of bone mass and a disruption of bone microarchitecture. Men lose bone density as they age, resulting in brittle bones. The loss of free testosterone is one of the key factors. The objective of present study was to evaluate Allolobophora caliginosa extract (AcE) for its anti-osteoporotic and antiapoptotic activity in orchiotomized rat model at two different dose levels. Twenty eight male rats were divided into two groups. The first group represented sham operated rats while the second group underwent bilateral orchidectomy (OCX). After one week of recovery from orchidectomy surgery, the second group was randomly subdivided into 3 subgroups. The first OCX subgroup was administered orally distilled water daily for 10 weeks. The other two OCX subgroups were administered AcE (100 or200 mg/kg body weight/day) orally for 10 weeks. Orchiectomy induces remarkable loss of the cortical as well as trabecular bone loss; which, could be counterbalanced by Allolobophora caliginosa extract (AcE) that prevented cortical as well as trabecular bone loss. Allolobophora caliginosa extract (AcE) at Dose 200 mg/kg/day was found to be effective at a highly significant level in osteoporotic bone, as determined by histological images and immunohistochemical study, where Dose (100 mg/kg/day) was found to be moderately significant.In the present study, it is suggested that AcE may inhibit steroid-induced osteoblasts apoptosis, potentially via upregulation of Bcl-2 and downregulation of caspase-3. Allolobophora caliginosa extract demonstrates anti-apoptotic and anti-oxidant properties. Therefore, AcE may be used for the prevention of steroid-induced bone damage.

RESUMEN: La osteoporosis es una afección ósea caracterizada por una pérdida de masa ósea y una alteración de la microarquitectura ósea. Los hombres pierden densidad ósea a medida que envejecen, lo que resulta en huesos quebradizos. La pérdida de testosterona libre es factor clave en este proceso. El objetivo del presente estudio fue evaluar el extracto de Allolobophora caliginosa (AcE) debido a su actividad antiosteoporótica y antiapoptótica en un modelo de rata orquiectomizadas con dos niveles de dosis diferentes. Se dividieron veintiocho ratas macho en dos grupos. El primer grupo incluyó ratas con operación simulada, mientras que el segundo grupo se sometió a orquidectomía bilateral (OCX). Después de una semana de recuperación de la orquidectomía, el segundo grupo fue subdividido en 3 subgrupos. Al primer subgrupo de OCX se administró diariamente agua destilada por vía oral durante 10 semanas. Los otros dos subgrupos de OCX se administraron por vía oral AcE (100 o 200 mg / kg de peso corporal / día) durante 10 semanas. La orquidectomía induce una pérdida notable del hueso cortical y trabecular; el cual podría ser contrarrestado por el extracto de Allolobophora caliginosa (AcE) que previno la pérdida de hueso tanto cortical como trabecular visualizado en imágenes histológicas y estudio inmuno- histoquímico, donde se encontró que la dosis (100 mg / kg / día) era moderadamente significativa. En el presente estudio, se sugiere que la AcE puede inhibir la apoptosis de los osteoblastos inducida por esteroides, potencialmente a través de la regulación al alza de Bcl 2 y la regulación a la baja de caspasa 3. El extracto de Allolobophora caliginosa demuestra propiedades anti apoptóticas y antioxidantes. Por lo tanto, AcE puede usarse para la prevención del daño óseo inducido por esteroides.

Preparation of purified proteins from fresh Pheretima and their inhibitory effect against pulmonary fibrosis in mice / 南方医科大学学报

OBJECTIVE@#To develop a convenient method for rapid purification of fresh Pheretima proteins and assess the inhibitory effect of these proteins against pulmonary fibrosis.@*METHODS@#The crude extract of fresh Pheretima was obtained by freeze-drying method and then purified by size exclusion chromatography. The composition of the purified proteins was analyzed by mass spectrometry. MRC-5 cells were treated with 5 ng/mL TGF-β1 alone (model group) or in combination with SB431542 (2 μmol/L) or the purified proteins (13.125 μg/mL), and the cytotoxicity of purified proteins and their inhibitory effects on cell proliferation were detected with CCK8 assay. Flow cytometry was used to detect the changes in cell apoptosis, and the cellular expressions of α-SMA, Vimentin, E-cadherin, collagen I, Smad2/3 and P-Smad2/3 were detected using RT-PCR and Western blotting. In the animal experiment, adult male C57BL/6 mice were subjected to intratracheal instillation of bleomycin followed by treatment with the purified proteins (5 mg/mL) for 21 days, after which HE and Masson staining was used to observe the pathological changes in the lung tissue of the mice.@*RESULTS@#We successfully obtained purified proteins from fresh Pheretima protein by size exclusion chromatography. Treatment with the purified proteins significantly inhibited TGF-β1-induced proliferation of MRC-5 cells (P < 0.01), reduced the cellular expressions of α-SMA, Vimentin and collagen I (P < 0.001 or P < 0.01), increased the expression of E-cadherin (P < 0.01), and inhibited the expressions of Smad2/3 and P-Smad2/3 (P < 0.001 or P < 0.01). In male C57BL/6 mice models of bleomycin-induced pulmonary fibrosis, treatment with the purified proteins obviously reduced the number of inflammatory cells and fibrotic area in the lungs.@*CONCLUSION@#The purified proteins from fresh Pheretima obtained by size exclusion chromatography can inhibit pulmonary fibrosis in mice by regulating the TGF-β/ Smad pathway.

Research progress on Pheretima and earthworms with related origin / 中国中药杂志

Through literature analysis of Pheretima and its origin-related earthworm,this study summarized the progress on Pheretima in textual criticism of origin,origin identification,effective components,detection of harmful components,and pharmacological effects,which can lay a basis for further research on Pheretima. Through literature research,the authors found that Pheretima was first recorded in Secret Formulary for Traumatology and Fracture Taught by Immortal written by LIN Daoren in Tang Dynasty rather than the Taiping Holy Prescriptions for Universal Relief in Song Dynasty. The latest techniques for origin identification include microscopic trait identification,DNA barcoding,and HPLC. The main effective components of Pheretima are proteins,polypeptides,enzymes,nucleotides,amino acids,and trace elements. According to recent studies,Pheretima has anti-pulmonary and anti-renal interstitial fibrosis,respiratory syncytial virus-inhibiting,human hypertrophic scar fibroblast proliferation-suppressing,and mouse embryonic fibroblast proliferation-promoting effects. Moreover,Pheretima can prevent colitis-induced colon cancer by inhibiting the activation of COX-2/PGE2/β-catenin signaling pathway. METHODS:: for detecting the harmful components and their residues( organic pollutant polychlorinated biphenyl,heavy metals) and bacteria in Pheretima,have been established. Pheretima,mainly derived from wild earthworms,has remarkable clinical efficacy. However,the wild resource is in short supply and artificial culture is expected to be a promising solution.

Edaphic macrofauna in degradation of animal and vegetable residues / Macrofauna edáfica no processo de vermicompostagem de resíduos animais e vegetais

Abstract The objective of this study was to evaluate the ability to degrade organic matter by edaphic macrofauna (worms), carbon/nitrogen (C/N) ratio and hydrogenation potential (pH) during the vermicomposting process in different organic residues. The treatments were constituted by organic residues of animal origin (bovine, ovine and equine manure) and vegetable (herb-checkmate and coffee drag), which were conditioned in plastic pots with a capacity of 10 liters, comprising five treatments in a completely randomized experimental design, with five replications. Were inoculated 150 earthworms of the species Eisenia foetida, into each plot. After 87 days, the evaluation of the multiplication of the earthworms was carried out, through its manual count and its cocoons. At the beginning and at the end of the experiment, the samples were submitted to analysis of humidity at 60 °C, pH, volumetric density, chemical analysis of macronutrients and C/N ratio. There was a dominance of worms and cocoons in the process of vermicomposting in the residues of ovine manure and herb-checkmate. The macronutrients (P, K and Mg) and C/N ratio were higher in the vegetal residues, while for N higher values were found in ovine manure and coffee drag treatments, and for Ca higher value among treatments was observed in the coffee drag treatment at the end and the lowest value at initiation. The results obtained in this study demonstrate the importance of the edaphic macrofauna to the vermicomposting process, since it allows more information about its influence on the continuity of soil organic matter decomposition processes.

Resumo Este estudo objetivou avaliar a capacidade de degradar a matéria orgânica pela macrofauna edáfica (minhocas), a relação carbono/nitrogênio (C/N) e o potencial hidrogeniônico (pH), durante o processo da vermicompostagem em diferentes resíduos orgânicos. Os tratamentos foram constituídos por resíduos orgânicos de origem animal (esterco bovino, ovino e equino) e vegetal (resíduo de erva-mate e borra-de-café), os quais foram acondicionados em vasos plásticos com capacidade de 10 litros, compondo cinco tratamentos em delineamento experimental inteiramente casualisado, com cinco repetições. Foram inoculadas 150 minhocas da espécie Eisenia foetida, em cada recipiente. Após 87 dias, foi realizada a avaliação da multiplicação das minhocas, através da sua contagem manual e seus casulos. Os resíduos foram submetidos, ao inicio e ao final do experimento, a análises de umidade a 60 °C, pH, densidade volumétrica, análise química de macronutrientes e relação C/N. Houve uma dominância de minhocas e casulos no processo da vermicompostagem nos resíduos de esterco ovino e erva-mate. Observou-se para os macronutrientes (P, K e Mg) e para a relação C/N uma maior quantidade nos resíduos vegetais, enquanto que para N valores maiores foram encontrados nos tratamentos esterco ovino e borra-de-café, e para Ca o maior valor entre os tratamentos foi observado no tratamento com borra de café ao final e o menor valor no inicio. Os resultados obtidos neste estudo demonstram a importância da macrofauna edáfica para o processo da vermicompostagem por possibilitar maiores informações sobre sua influência na continuidade dos processos de decomposição da matéria orgânica.

Study on proteins in Guangdilong by nano LC/orbitrap fusion lumos HR-MS / 中国中药杂志

In the present study,fresh Guangdilong( GD),originating from Pheretima aspergillum,was taken as the object. The total proteins from GD were firstly separated by SDS-PAGE according to their molecular weights and in-gel digestion was then performed.After that,the peptides were analyzed by nano LC/orbitrap fusion lumos high resolution mass spectrometry( nano LC/orbitrap fusion lumos HR-MS). Protein identification was implemented by comparison with Annelida. fasta database using Proteome Discoverer software.As a result,386 proteins were tentatively identified,including chain F,globin B chain,glyceraldehyde-3-phosphate dehydrogenase,fibrinolytic protein,and so on. Most of the proteins took part in cell structure and energy metabolism,and fibrinolytic protein and lombricine kinase might be related to fibrinolytic activity. Protein classification based on gene ontology was carried out using PANTHER and KEGG for metabolic pathway enrichment. The results indicated that these proteins were related to diverse signal transduction pathways,including metabolic pathways,central carbon metabolism,biosynthesis of amino acids,ribosome,glycolysis,citrate cycle( TCA cycle),and so on. This study would lay the foundation for the further research on the proteins in GD and also their functions.

The Connection between Charles Darwin's Evolutionary Theory of ‘Heredity of Behaviors’ and the 19th Century Neuroscience: The Influence of Neuroscience on Darwin's Overcoming of Lamarck's Theory of Evolution / 의사학

The nineteenth century neuroscience studied the instinct of animal to understand the human mind. In particular, it has been found that the inheritance of unconscious behavior like instinct is mediated through ganglion chains, such as the spinal cord or sympathetic nervous system, which control unconscious reflexes. At the same time, the theory of Inheritance of Acquired Characteristics (hereafter ‘IAC’) widely known as Lamarck's evolutionary theory provided the theoretical frame on the origin of instinct and the heredity of action that the parental generation's habits were converted into the nature of the offspring generation. Contrary to conventional knowledge, this theory was not originally invented by Lamarck, and Darwin also did not discard this theory even after discovering the theory of natural selection in 1838 and maintained it throughout his intellectual life. Above all, in the field of epigenetics, the theory of ‘IAC’ has gained attention as a reliable scientific theory today. Darwin discovered crucial errors in the late 1830s that the Lamarck version's theory of ‘IAC’ did not adequately account for the principle of the inheritance of unconscious behavior like instinct. Lamarck's theory regarded habits as conscious and willful acts and saw that those habits are transmitted through the brain to control conscious actions. Lamarck's theory could not account for the complex and elaborate instincts of invertebrate animals, such as brainless ants. Contrary to Lamarck's view, Darwin established the new theory of ‘IAC’ that could be combined with contemporary neurological theory, which explains the heredity of unconscious behavior. Based on the knowledge of neurology, Darwin was able to translate the ‘principle of habit’ into a neurological term called ‘principle of reflex’. This article focuses on how Darwin join the theory of ‘IAC’ with nineteenth century neuroscience and how the neurological knowledge from the nineteenth century contributed to Darwin's overcoming of Lamarck's ‘IAC’. The significance of this study is to elucidate Darwin's notion of ‘IAC’ theory rather than natural selection theory as a principle of heredity of behavior. The theory of ‘IAC’ was able to account for the rapid variation of instincts in a relatively short period of time, unlike natural selection, which operates slowly in geological time spans of tens of millions of years. The nineteenth century neurological theory also provided neurological principles for ‘plasticity of instinct,’ empirically supporting the fact that all nervous systems responsible for reflexes respond sensitively to very fine stimuli. However, researchers of neo-Darwinian tendencies, such as Richard Dawkins and evolutionary psychologists advocating the ‘selfish gene’ hypothesis, which today claim to be Darwin's descendants, are characterized by human nature embedded in biological information, such as the brain and genes, so that it cannot change at all. This study aims to contribute to reconstructing the evolutionary discourse by illuminating Darwin's insights into the “plasticity of nature” that instincts can change relatively easily even at the level of invertebrates such as earthworms.

Development and multiplication of Eisinea andrei in the manure of cattle subjected to high Trichoderma doses / Desenvolvimento e multiplicação de Eisenia andrei em esterco bovino em altas doses de Trichoderma

The efficacy of the vermicomposting and products based on the antagonistic fungus and plant growth promoter trichoderma (Trichoderma spp) is well known and studied in organic agriculture. However, for a better methodological efficiency are necessary studies to evaluate the effect of high doses of these bioproducts in the biology and development of earthworms. Thus, the present work aims to test the use of high commercial biocontrol product (ICB Nutrisolo Trichoderma) doses by evaluating the multiplication and development of Eisenia andrei. Changes in the chemical features of the substrate produced by the vermicomposting process using in natura and sterilized organic cattle manure were also assessed. Each experimental unit consisted of 6 kg of substrate (in multipurpose polypropylene box ­ 20 x 40 x 50 cm) containing 48 clitelate adult Eisenia andrei earthworms. ICB Nutrisolo Trichoderma was used as biological agent along with eight strains of the following species: T. koningiopsis, T. asperellum and T. harzianum. The following treatments were applied at doses of 1011 CFU kg-1 of ICB Nutrisolo Trichoderma in the presence of earthworms: T1 (0.5); T2 (1.0); T3 (2.0); T4 (4.0); T5 (8.0) and T6 (0.0). The T7 treatment was herein used in order to evaluate the chemical features of the vermicompost. It was a completely randomized design with four replications per treatment. The temperature was kept at 28°C and humidity ranged between 60 and 70%. After 60 days, the number of young and adult earthworms, and cocoons was counted; then, their dry biomass was assessed. The results found in the lethality test showed decrease in the number of earthworms treated with 4.0x1011 CFU kg-1 of ICB. The biological product doses up to 1.0x1011 CFU kg-1 did not alter the number of adult earthworms and cocoons, or the multiplication index of E. andrei in cattle waste vermicomposts. There was no influence of the tested doses on earthworms' individual development. However, doses above 2.0x1011 CFU kg-1 decreased their total biomass. The C/N ratio for all treatments indicates maturity within acceptable results for organic compounds.

A eficácia da vermicompostagem e de bioprodutos à base do fungo antagonista e promotor de crescimento vegetal trichoderma (Trichoderma spp) é bem conhecida e estudada na agricultura orgânica. Entretanto, para uma melhor eficiência metodológica, são necessários estudos que possam avaliar a interferência de altas doses desses bioprodutos na biologia e desenvolvimento das minhocas. Baseado nesse contexto, o objetivo deste trabalho foi testar altas doses do produto comercial biológico ICB Nutrisolo Trichoderma (ICB), avaliando-se a multiplicação e desenvolvimento de Eisenia andrei, bem como alterações nas características químicas do substrato produzido no processo de vermicompostagem, a partir do resíduo orgânico esterco bovino. O esterco bovino in natura foi autoclavado a 121°C, por duas vezes, em um intervalo de 24 h. A unidade experimental constituiu-se de 6 kg de substrato condicionados em caixa multiuso de polipropileno, com dimensões 20 x 40 x 50 cm, contendo 48 minhocas adultas e cliteladas da espécie E. andrei. Como agente biológico, utilizou-se o produto comercial ICB na forma de fluído, composto por oito cepas das espécies T. koningiopsis, T. asperellum e T. harzianum, com as seguintes doses nos tratamentos a seguir: T1 (0.5); T2 (1.0); T3 (2.0); T4 (4.0); T5 (8.0); e T6 (0.0), sendo todas as concentrações em 1011 UFC kg-1 do produto em vermicomposto, e, para a avaliação das características químicas do vermicomposto em altas doses do produto ICB, foi utilizado também o T7 (somente substrato). O delineamento foi inteiramente casualizado com quatro repetições por tratamento. A temperatura foi mantida a 28ºC e a umidade entre 60 e 70%. Após 60 dias do início da instalação, fez-se a contagem do número de minhocas adultas, jovens e casulos; posteriormente, avaliou-se o seu peso seco total. Os resultados observados no teste de letalidade mostram que, somente a partir de 4.0x1011 UFC kg-1 de ICB, há decréscimo do número de minhocas. Doses altas até 1.0x1011 UFC kg-1 do produto não alteram o número de minhocas adultas e de casulos de E. andrei em vermicompostagem com esterco bovino, entretanto, o índice de multiplicação foi inferior em todos os tratamentos com o produto. Doses acima de 2.0x1011 UFC kg-1 diminuíram o peso seco total. A relação C/N em todos os tratamentos indica maturidade dentro de resultados aceitáveis para compostos orgânicos.

Current knowledge of earthworm richness and distribution in Santa Fe province, Argentina / Conocimiento actual sobre riqueza y distribución de lombrices de tierra en la provincia de Santa Fe, Argentina

Abstract Updated list of earthworms (Oligochaeta: Lumbricina) from Santa Fe (Argentina) is presented in this work, including current data of species richness and territorial distribution status and information collected by Ljungström and collaborators 40 years ago. Field samplings were conducted between 2012 and 2015 at 23 sites, located in 11 of the 19 districts of the province. Earthworms were collected following a standard methodology. The conservation of specimens was done with 4 % formalin solution and their identification was performed according to taxonomy keys. Richness, similarity and complementarity of species between the phytogeographic provinces were analyzed. A total of 15 earthworm species were identified and grouped into ten genera and five families: Acanthodrilidae (Dichogaster bolaui, Microscolex dubius), Glossoscolecidae (Glossodrilus parecis), Lumbricidae (Aporrectodea caliginosa, Aporrectodea rosea, Aporrectodea trapezoides, Bimastos parvus, Eisenia fetida, Octolasion tyrtaeum), Megascolecidae (Amynthas gracilis, Amynthas morrisi, Metaphire californica), Ocnerodrilidae (Eukerria saltensis, Eukerria rosae, Eukerria stagnalis). From all the species found five, G. parecis, E. saltensis, E. rosea, E. stagnalis and M. dubius, are native to South America, and the rest were introduced from Asia and Europe. The Espinal presented the greatest richness of earthworms (12), while the Pampeana showed high values of complementarity (greater than 70 %) with the Chaco Húmedo and Valle de Inundación del Río Paraná. Endogeic species were present in all environments surveyed. In particular, the exotic species A. trapezoides and A. morrisi showed a wide geographical distribution, having been found at 70 and 50 %, respectively, of the total number of the studied sites. One species, E. rosea, which is in the list, was not recorded in the sampling of 40 years ago. The results of current survey show that the number of species was lower compared to study by Ljungström and collaborators (60 % of the 23 species registered). Possibly the remarkable change in the richness and distribution of earthworms could be a process associated with changes in soil use and land management developed over 40 years. Rev. Biol. Trop. 66(3): 1171-1181. Epub 2018 September 01.

Resumen En este trabajo se presenta una lista actualizada de lombrices de tierra (Oligochaeta: Lumbricina) de Santa Fe (Argentina), incluyendo datos actuales de riqueza de especies y estado de distribución territorial e información recolectada por Ljungström y colaboradores hace 40 años. Los muestreos de campo se realizaron entre 2012 y 2015 en 23 sitios, ubicados en 11 de los 19 departamentos de la provincia. Las lombrices fueron recolectadas siguiendo métodos estándar. La conservación de los especímenes se realizó con solución de formalina al 4 % y la identificación mediante claves taxonómicas. La riqueza, similitud y complementariedad de especies, se analizó entre las provincias fitogeográficas y se identificaron un total de 15 especies de lombrices en diez géneros y cinco familias: Acanthodrilidae (Dichogaster bolaui, Microscolex dubius), Glossoscolecidae (Glossodrilus parecis), Lumbricidae (Aporrectodea caliginosa, Aporrectodea rosea, Aporrectodea trapezoides, Bimastos parvus, Eisenia fetida, Octolasion tyrtaeum), Megascolecidae (Amynthas gracilis, Amynthas morrisi, Metaphire californica), Ocnerodrilidae (Eukerria saltensis, Eukerria rosae, Eukerria stagnalis). De todas las especies encontradas, cinco: G. parecis, E. saltensis, E. rosea, E. stagnalis y M. dubius, son nativas de América del Sur, y el resto introducidas desde Asia y Europa. Las especies endógeas estuvieron presentes en todos los ambientes estudiados. En particular, las especies exóticas: A. trapezoides y A. morrisi, mostraron amplia distribución geográfica, se encuentran en el 70 y 50 %, respectivamente, de los sitios estudiados. Una especie, E. rosea, que está en la lista, no fue registrada en el muestreo de hace 40 años. La zona del Espinal presentó la mayor riqueza de lombrices (12), en tanto que la Pampeana mostró valores altos de complementariedad (mayores al 70 %) con el Chaco Húmedo y el Valle de Inundación del Río Paraná. Los resultados del relevamiento actual muestran que el número de especies fue menor en comparación con el estudio de Ljungström y colaboradores (60 % de las 23 especies registradas). Posiblemente el notable cambio en la biodiversidad del paisaje de la provincia, asociado al uso de la tierra y a la gestión de los sistemas de producción desarrollados a lo largo de 40 años, podría haber influido en la distribución y riqueza de especies de lombrices.

Avaliação da atividade da enzima glutationa-s-transferase da minhoca Eisenia foetida exposta à ivermectina / Evaluation of glutatione-s-transferase enzyme activity of the earthworm Eisenia foetida exposed to ivermectin

Muitos tipos de drogas são usados na medicina veterinária para controlar e melhorar a saúde animal através de tratamentos terapêuticos e profiláticos. A desvantagem desta prática é que os produtos farmacêuticos e seus metabólitos são liberados no ambiente e podem influenciar a fauna do solo através da excreção do esterco ou pela posterior aplicação ao campo agrícola. As avermectinas são vastamente utilizadas na medicina veterinária e na agricultura. Estudos anteriores demonstraram que a ivermectina (IVM), um parasiticida amplamente utilizado, é muito tóxico para diversas espécies de invertebrados não-alvo. Tendo em vista que a IVM é pouco metabolizada, excretada de forma relativamente inalterada e pela escassez de dados sobre a toxicidade aos invertebrados do solo, foram investigados, neste estudo, os efeitos agudos e crônicos deste parasiticida sobre a glutationa-s -transferase (GST) da oligoqueta Eisenia foetida. As minhocas Eisenia foetida foram expostas à concentrações de IVM a 0, 1, 5, 10, 50 e 100 mg kg-1, e as amostras foram tomadas nos dias 7, 14 e 28 para determinação da atividade da GST. Os resultados mostraram que a duração da exposição alterou significativamente os efeitos do parasiticida investigado sobre a atividade de GST. Especificamente, após uma redução inicial, o prolongamento da exposição causou a indução da atividade da GST. Com o aumento da concentração de IVM, as atividades da GST foram inibidas significativamente após 7 dias de exposição. Em particular, o efeito inibitório foi significativo nas concentrações mais elevadas de tratamento (10, 50 e 100 mg kg-1). Por outro lado, aos 14 e 28 dias foram observadas induções na atividade da enzima. A atividade da GST pode ser considerada como parâmetro sensível para avaliar a toxicidade da ivermectina para minhocas.

Many types of drugs are used in veterinary medicine to control and improve animal health through therapeutic and prophylactic treatments. The disadvantage of this practice is that pharmaceuticals and their metabolites are released into the environment and may influence soil fauna through manure excretion and subsequent application to agricultural field. The avermectins are extensively and increasingly used in veterinary medicine and agriculture. Previous studies have shown that ivermectin (IVM), a widely used parasiticide, is very toxic to many non-target invertebrate species. In view of the little metabolism and most of the ivermectin dose given to the animal is excreted, relatively unaltered, primarily in the feces and the scarcity of data on toxicity to soil invertebrates, acute and chronic effects of the parasiticide on the glutatione-s-transferase (GST) of the oligochaete Eisenia foetida were investigated. Earthworms of Eisenia foetida were exposed to IVM at 0, 1, 5, 10, 50 and 100 mg kg-1 concentrations; samples were taken at days 7, 14, and 28 exposure for determination of GST activities. The results showed that duration of the exposure significantly changed the effects of the investigated parasiticide on the GST activity. Namely, after the initial decrease, the prolongation of exposure caused the induction of the GST activity. With increasing IVM concentration, GST activities were inhibited significantly after 7 days of the exposure. In particular, the inhibition effect was significant at the higher treatment levels (10, 50 and 100 mg kg-1). On the other hand, at 14 and 28 days were observed inductions of enzyme activity. GST activity can be regarded as sensitive parameter for evaluating the toxicity of ivermectin to earthworms.

Presence of Eisenia fetida enhanced phytoremediation of anthracene by Lolium perenne / Presença de Eisenia fetida aumentou a fitorremediação do antraceno por Lolium perene

The efficiency of co-application of Eisenia fetida and ryegrass was evaluated in a process called earthworm-assisted phytoremediation. Anthracene was used as a model compound for polycyclic aromatic hydrocarbons (PAHs). The experiments were conducted on a loamy soil in greenhouse conditions. At the end of the experiment, the soil samples were analyzed for residual anthracene by HPLC. Results showed that, phytoremediation using ryegrass could remove 81% of anthracene; however, the rate of removal was 92% when E. fetida was applied simultaneously. E. fetida alone could also remove the initial concentration of anthracene by 40%. Although ryegrass itself could remove anthracene significantly, the employment of earthworm, together with plant was more efficient than each of them individually. The application of E. fetida could also enhance the growth parameters of ryegrass significantly. In comparison to the control, the presence of E. fetida increased plant dry weight (7.8%), root length (47%), shoots length (32%), and root volume (12%). The number of live earthworms was also increased in the planted pots, indicating the helpfulness of the plant for survival of the earthworm in the PAH-contaminated soil. Although plant and earthworm use completely different mechanisms for anthracene degradation, they improve efficiency and survival of the three-component-system.

A eficiência da co-aplicação de Eisenia fetida e azevém foi avaliada em um processo denominado fitorremediação assistida por minhocas. O antraceno foi usado como um composto modelo para hidrocarbonetos aromáticos policíclicos (PAHs). Os experimentos foram conduzidos em um solo argiloso em condições de estufa. No final da experiência, as amostras de solo foram analisadas quanto ao antraceno residual por HPLC. Os resultados mostraram que, a fitorremediação com azevém pode remover 81% do antraceno; no entanto, a taxa de remoção foi de 92% quando E. fetida foi aplicada simultaneamente. E. fetida sozinha também foi capaz de remover a concentração inicial de antraceno em 40%. Embora o próprio azevém pudesse remover significativamente o antraceno, o emprego da minhoca, juntamente com a planta, foi mais eficiente do que cada um deles individualmente. A aplicação de E. fetida também pode melhorar significativamente os parâmetros de crescimento do azevém. Em comparação com o controle, a presença de E. fetida aumentou o peso seco da planta (7,8%), o comprimento da raiz (47%), o comprimento da parte aérea (32%) e o volume radicular (12%). O número de minhocas vivas também aumentou nos vasos plantados, indicando a utilidade da planta para a sobrevivência da minhoca no solo contaminado com PAH. Embora plantas e minhocas usem mecanismos completamente diferentes para a degradação do antraceno, eles melhoram a eficiência e a sobrevivência do sistema de três componentes.

Organophosphorous pesticides: their effects on biosentinel species and humans: control and application in Chile / Pesticidas organofosforados: sus efectos en especies biosentinelas y humanos: control y aplicación en Chile

The environment is negatively affected by the increasing accumulation of both natural and man-made waste and by-products. Organophosphorous pesticides -malathion, diazinon and methamidophos- are used worldwide in pest control. The aim of this report is to review the effects of organophosphates on the male reproductive tract of mice, rats and earthworms, and to evaluate their projection into the human population. Assessing failures in the male reproductive system is an excellent in vivo biomarker to determine the level of toxicity of suspected pollutants. In rodents organophosphates cause decreased testicular weight and sperm density, abnormal tubular plugging and increased teratozoospermia. In earthworms they cause a significant decrease in body weight and alter the spermatheca, with an initial significant increase in immature sperm followed by a significant decrease in sperm count with high frequency of metachromasia. Given the environmental impact of these pesticides -and their potential effects on human health-, international and regional organizations are warning about the correct handling and managing of these substances during work-related and domestic exposures and about their relation to water sources and food, placing a greater emphasis on the school children population due to its higher vulnerability, reduced detoxification capacity, and their endocrine and physiological effects.

El medio ambiente se ve afectado negativamente por la creciente acumulación de desechos y subproductos naturales y artificiales. Los plaguicidas organofosforados malatión, diazinón y metamidofos, son usados en todo el mundo en el control de plagas. El objetivo de este trabajo fue revisar los efectos de los organofosforados en el tracto reproductivo masculino de ratones, ratas y lombrices de tierra, y evaluar su proyección en la población humana. La evaluación de fallas en el sistema reproductor masculino es un excelente biomarcador in vivo para determinar el nivel de toxicidad de los contaminantes químicos. En roedores, los organofosforados causan disminución del peso testicular y de la densidad espermática, obstrucción tubular anormal y aumento de la teratozoospermia. En lombrices de tierra causan una disminución significativa en el peso corporal y alteran la espermateca, con un aumento inicial significativo en espermatozoides inmaduros, seguido de una disminución significativa en el recuento de espermatozoides con alta frecuencia de metacromasia. Dado el impacto medioambiental de estos plaguicidas y sus efectos potenciales en la salud humana, las organizaciones internacionales y regionales advierten sobre el manejo y uso correctos de estas sustancias durante exposiciones laborales y domésticas y sobre su relación con la contaminación de las fuentes de agua y alimentos, colocando énfasis en la población de niños en edad escolar, debido a su mayor vulnerabilidad, reducción de la capacidad de desintoxicación y sus efectos a nivel endocrino y fisiológico.

De novo transcriptome assembly, functional annotation and differential gene expression analysis of juvenile and adult E. fetida, a model oligochaete used in ecotoxicological studies

BACKGROUND: Earthworms are sensitive to toxic chemicals present in the soil and so are useful indicator organisms for soil health. Eisenia fetida are commonly used in ecotoxicological studies; therefore the assembly of a baseline transcriptome is important for subsequent analyses exploring the impact of toxin exposure on genome wide gene expression. RESULTS: This paper reports on the de novo transcriptome assembly of E. fetida using Trinity, a freely available software tool. Trinotate was used to carry out functional annotation of the Trinity generated transcriptome file and the transdecoder generated peptide sequence file along with BLASTX, BLASTP and HMMER searches and were loaded into a Sqlite3 database. To identify differentially expressed transcripts; each of the original sequence files were aligned to the de novo assembled transcriptome using Bowtie and then RSEM was used to estimate expression values based on the alignment. EdgeR was used to calculate differential expression between the two conditions, with an FDR corrected P value cut off of 0.001, this returned six significantly differentially expressed genes. Initial BLASTX hits of these putative genes included hits with annelid ferritin and lysozyme proteins, as well as fungal NADH cytochrome b5 reductase and senescence associated proteins. At a cut off of P = 0.01 there were a further 26 differentially expressed genes. CONCLUSION: These data have been made publicly available, and to our knowledge represent the most comprehensive available transcriptome for E. fetida assembled from RNA sequencing data. This provides important groundwork for subsequent ecotoxicogenomic studies exploring the impact of the environment on global gene expression in E. fetida and other earthworm species.

Assessment of benthic macroinvertebrates at Nile tilapia production using artificial substrate samplers / Avaliação da comunidade de macroinvertebrados bentônicos em produção de tilápia-do-Nilo com uso de coletores com substrato artificial

Abstract Biomonitoring is a cheap and effective tool for evaluation of water quality, and infer on the balance of aquatic ecosystems. The benthic macroinvertebrates are bioindicators sensitive to environmental changes, and can assist in detecting and preventing impacts such as organic enrichment and imbalance in the food chain. We compared the structure of benthic communities on artificial substrate samplers located in places near and far from net cages for production of Nile tilapia (Oreochromis niloticus). Samplers were manufactured with nylon net, using substrates such as crushed stone, gravel, loofah and cattail leaves. Samples were collected after 30 days of colonization, rinsed and then the specimens were identified and quantified. The following metrics were calculated: richness of Operational Taxonomic Units, Margalef richness, abundance of individuals, Shannon index and evenness index. The macrobenthic community structure was strongly modified according to the proximity of the net cages. Metrics showed significant differences (p < 0.05) between near and distant sites, for both periods (dry and rainy seasons). The position of the samplers significantly affected the structure of macroinvertebrate community, as near sites showed higher values for the community metrics, such as richness and diversity. Near sites presented a larger number of individuals, observed both in the dry and rainy seasons, with a predominance of Chironomidae (Diptera) in the dry season and Tubificidae (Oligochaeta) in the rainy season.

Resumo O biomonitoramento é uma ferramenta com custo relativamente baixo e eficaz para avaliação da qualidade da água, além de inferir sobre o equilíbrio dos ecossistemas aquáticos. Os macroinvertebrados bentônicos são bioindicadores sensíveis às mudanças ambientais, e podem auxiliar na detecção e prevenção de impactos como o enriquecimento orgânico e o desequilíbrio da cadeia alimentar. Como objetivo, foi comparada a estrutura das comunidades bentônicas em coletores com substrato artificial situados em locais próximos e distantes de tanques rede para produção de tilápia do Nilo (Oreochromis niloticus). Os coletores foram fabricados com rede de náilon, utilizando os substratos brita, cascalho, bucha vegetal e folhas de taboa. Foram realizadas coletas após 30 dias de colonização. As amostras foram lavadas e os espécimes identificados e quantificados. As métricas calculadas foram: riqueza de Unidades Taxonômicas Operacionais, riqueza de Margalef, abundância de indivíduos, índice de Shannon e índice de Pielou. A estrutura da comunidade macrobentônica foi fortemente modificada de acordo com a proximidade dos tanques-rede. As métricas apresentaram diferenças significativas (p < 0,05) entre os locais próximos e distantes, em ambos os períodos (estações seca e chuvosa). A posição dos amostradores afetou significativamente a estrutura da comunidade de macroinvertebrados, na medida em que locais próximos dos tanques rede apresentaram valores mais elevados para as métricas de comunidade, como riqueza e diversidade. Amostras próximas aos tanques rede apresentaram um maior número de indivíduos em ambas as estações do ano, com predomínio de Chironomidae (Diptera) na estação seca e Tubificidae (Oligochaeta) na estação chuvosa.

Glutathione-S-transferase production in earthworm (Annelida: Eudrilidae) as a tool for heavy metal pollution assessment in abattoir soil

AbstractThe use of direct response of animals to environmental challenges by production of biomarkers is a better tool to assess environmental pollution than the conventional methods. This study aimed to measure Glutathione-S-transferase (GST) in earthworms as tools for assessing heavy metal pollution in abattoir soil. Five (5) replicates each of earthworm species (Libyodrilus violaceous, Eudrilus eugeniae and Alma millsoni), soil and rumen waste samples were collected from three (3) abattoir sites (Lafenwa, Gbonogun and Madojutimi abattoirs), and a control site located within Federal University of Agriculture Abeokuta, beside an undisturbed stream with no rumen waste. Heavy metal (Cu, Zn, Pb, Cd, Co, Cr, Ni and Mn) concentrations in rumen waste, abattoir soils and earthworm tissues were determined using Atomic Absorption Spectrophotometer. The pH and organic matter (OM) concentrations of the rumen waste and abattoir soils were determined by standard methods. GST activities in the earthworm tissues were determined through the conjugation of 1 mM reduced glutathione (GSH) with 1 mM 1-chloro-2,4-dinitrobenzene (CDNB). The rumen waste recorded significantly higher (p ≤ 0.05) % OM, heavy metal concentrations and pH level than in their respective abattoir soils. The mean heavy metal concentrations of Cu, Zn, Pb, Cd and Mn were highest in the tissue of earthworm species obtained from Lafenwa abattoir. A significantly (p ≤ 0.05) higher GST activities were recorded in the tissue of earthworm species obtained from Lafenwa and Gbonogun abattoirs. Libyodrilous violaceus obtained from Lafenwa abattoir recorded the highest GST activity (8.47±1.39) in their tissue followed by the ones from Gbonogun abattoir (8.21±0.85). A significant (p ≤ 0.05) positive correlations was observed between GST activities in earthworm tissues and heavy metal concentrations. GST activities can therefore be used to assess the level of heavy metal pollution in abattoir soils. Rev. Biol. Trop. 64 (2): 779-789. Epub 2016 June 01.