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.

Microbial diversity and digestive enzyme activities in the gut of earthworms found in sawmill industries in Abeokuta, Nigeria / Diversidad microbiana y actividad de enzimas digestivas en el intestino de las lombrices de tierra en los aserraderos de Abeokuta, Nigeria

The growing demand for wood has resulted in large volumes of wood wastes that are daily released to the soil from the activities of sawmills in South-Western Nigeria. In an attempt to setup a bioremediation model for sawdust, this study therefore aimed at evaluating microbial diversity, and the level of digestive enzymes in the gut of earthworms (Eudrilus eugeniae, Libyodrilus violaceous and Hyperiodrilus africanus) of sawmill origin. Four major sawmills located in Abeokuta (7o9’12” N - 3o19’35” E), namely Lafenwa, Sapon, Isale-Ake and Kotopo sawmills were used for this study. The arboretum of the Federal University of Agriculture, Abeokuta was used as control. Gut microbial analysis was carried out using the pour-plate method while digestive enzyme activities in the earthworm guts were done by the spectrophotometric method. Higher microbial counts (28.5±0.1x10³-97.0±0.1x10³cfu for bacteria and 7.0±0.1x10³-96.0±0.1x10³cfu for fungi) and microbial diversity were recorded in the gut of earthworms of the sawmill locations than those of the control site (17.5±0.1x10³cfu for bacteria and 4.5±0.1x10³cfu for fungi). Streptococcus mutans and Proteus spp.were common in the gut of E. eugeniae, and L. violaceous from the study sawmills, while Streptococcus mutans were also identified in H. africanus, but absent in the gut of E. eugeniae from the control site. Cellulase (48.67±0.02mg/g) and lipase (1.81±0.01mg/g) activities were significantly higher (p<0.05) in the gut of earthworms from the control site than those of the study sawmills. Furthermore, amylase (α and β) activity was highest in the gut of earthworms from the sawmills. Variations observed in the gut microbial and digestive enzyme activities of earthworms from the study sawmills as compared to the control site suggests that earthworms, especially E. eugeniae, could be a better organism for use as bioremediator of wood wastes. Rev. Biol. Trop. 62 (3): 1241-1249. Epub 2014 September 01.

La creciente demanda de madera ha resultado en grandes volúmenes de residuos de madera que se lanzan diariamente a la tierra, procedentes de las actividades de los aserraderos en el suroeste de Nigeria. Se realizó este estudio en un intento de configurar un modelo de biorremediación de aserrín, con el objetivo de evaluar la diversidad microbiana y el nivel de las enzimas digestivas en el intestino de las lombrices de tierra (Eudrilus eugeniae, Libyodrilus violáceo y Hyperiodrilus africanus) de origen aserradero. Para este estudio se utilizaron cuatro grandes aserraderos ubicados en Abeokuta (7°9’12 ‘’ N - 3°19’35 ‘’ E), a saber: Lafenwa, Sapon, Isale-Ake y Aserraderos Kotopo. El arboreto de la Universidad Federal de Agricultura, Abeokuta se utilizó como control. El análisis microbiano se llevó a cabo utilizando el método de vertido de placa, mientras que las actividades de enzimas digestivas en los intestinos de las lombrices se realizaron por el método espectrofotométrico. Los recuentos microbianos más altos fueron 28.5±97.0±0.1x10³-0.1x10³cfu para las bacterias y 7.0±0.1x10³-96.0±0.1x10³cfu para los hongos y la diversidad microbiana registrada en el intestino de las lombrices de tierra de los lugares de aserraderos y las del sitio de control fueron: 17.5±0.1x10³cfu para bacterias y 4.5±0.1x10³cfu para hongos. Streptococcus mutans y Proteus spp. eran comunes en el intestino de E. eugeniae, y L. violaceous de los aserraderos de estudio, mientras que también se identificaron Streptococcus mutans en H. africanus, que estuvieron ausentes en el intestino de E. eugeniae en el sitio de control. Actividades de celulasa (48.67±0.02mg/g) y lipasa (1.81±0.01mg/g) fueron significativamente mayores (p<0.05) en el intestino de las lombrices de tierra en el sitio de control que las de los aserraderos de estudio. Por otra parte, la actividad de la amilasa (α y β) fue mayor en el intestino de las lombrices de tierra de los aserraderos. Las variaciones observadas en la microbiota intestinal y la actividad de las enzimas digestivas de las lombrices de tierra de los aserraderos de estudio, en comparación con el sitio de control, sugieren que las lombrices de tierra, especialmente E. eugeniae, podrían ser un mejor organismo para su uso como biorremediador de residuos de madera.

Novel thrombolytic therapy discovered from traditional oriental medicine using the earthworm.

Since a few thousand years ago, the earthworm has been used as a drug for various diseases in China and the Far East. However, modern scientific pharmacological studies have not so far been performed. We extracted a very strong fibrinolytic enzyme from the earthworm, Lumbricus rubellus. This enzyme was heat-stable and displayed a very broad optimal pH range. Purification of the enzyme was performed and three partially purified fractions were obtained. These three fractions were further subdivided, and six purified fractions (F-I-0, 1, 2, F-II, and F-III-1,2) were finally obtained. Based on results of their enzymatic activities against various substrates, the fraction I enzymes are thought to represent chymotrypsin-like enzymes and the fraction III enzymes to represent trypsin-like enzymes. The fraction II enzyme appears to be neither a trypsin-nor chymotrypsin-like enzyme nor an elastase. We therefore designed trials for in vivo experiments on human volunteers. 120 mg of lyophilized earthworm powder was administered orally to 7 healthy volunteers (aged 28-52 years old) three times after meals every day for 17 days. Blood was withdrawn once a day before and at 1, 2, 3, 8, 11 and 17 days after commencing the administration. The fibrin degradation products (FDP) value, tissue plasminogen activator (t-PA) antigen level and t-PA activities were measured in the blood. Before the administration, the t-PA antigen level was 5.6 +/- 0.38 ng/ml, and it gradually increased until the 17th day. The FDP level was increased on the 1st and 2nd day after the administration, but had decreased and normalized by the 17th day. The fibrinolytic activities also tended to show an increase during the experiment. These results suggest that earthworm powder represents a possible oral thrombolytic agent. The earthworm enzyme may thus be applicable for treating patients with thalassemia.