scholarly journals Using of essential oils in the treatment of mice infected with Trypanosoma evansi

2014 ◽  
pp. 4109-4115 ◽  
Author(s):  
Matheus D Baldissera ◽  
Aleksandro S Da Silva ◽  
Camila B Oliveira ◽  
Rodrigo A Vaucher ◽  
Roberto CV Santos ◽  
...  
Keyword(s):  
Essential Oils ◽  
Positive Control ◽  
Negative Control ◽  
Trypanosoma Evansi ◽  
Trypanocidal Activity ◽  
Copaiba Oil ◽  
Group A ◽  
Group B

ABSTRACTObjective. This study aimed to test the effectiveness of copaiba, andiroba and aroeira essential oils for controlling trypanosomosis by Trypanosoma evansi with mice as experimental model. Materials and methods. Sixty-six mice were divided into eleven groups (A to L) with six animals each. Group A was the unique composed by healthy and uninfected animals (negative control). Animals in groups B to L were inoculated with 0.1 mL of blood containing 2.7 x 106 trypanosomes. Group B was used as a positive control without treatment. In experiment were tested copaiba (C, D and E), andiroba (F, G and H) and aroeira (I, J and L) oils at doses of 0.6, 0.8 and 1.0 mL kg-1 to infected mice (T. evansi). Results. These protocols did not provide curative efficacy; however, the mice treated with highest dose of copaiba showed a significant increase in the longevity when compared others groups. Conclusions. Previously in our studies, these essential oils have shown trypanocidal activity in vitro, but when they were tested in vivo in mice infected with T. evansi, this trypanocidal activity, or the curative effect was not found, being only able to prolong the lifespan of the animals treated with copaiba oil.

scholarly journals Trypanocidal activity of human plasma on Trypanosoma evansi in mice

2012 ◽  
Vol 21 (1) ◽  
pp. 55-59 ◽  
Author(s):  
Aleksandro Schafer Da Silva ◽  
Marcos Rafael Kroeker Duck ◽  
Vinicius da Rosa Fanfa ◽  
Mateus Anderson Otto ◽  
João Tomaz Schmitt Nunes ◽  
...  
Keyword(s):  
Human Plasma ◽  
Positive Control ◽  
Trypanosoma Evansi ◽  
Blood Type ◽  
Therapeutic Success ◽  
Trypanocidal Activity ◽  
Group A ◽  
Group B ◽  
Group D ◽  
Apolipoprotein L1

This study aimed to test an alternative protocol with human plasma to control Trypanosoma evansi infection in mice. Plasma from an apparently 27-year-old healthy male, blood type A+, was used in the study. A concentration of 100 mg.dL-1 apolipoprotein L1 (APOL1) was detected in the plasma. Forty mice were divided into four groups with 10 animals each. Group A comprised uninfected animals. Mice from groups B, C and D were inoculated with a T. evansi isolate. Group B was used as a positive control. At three days post-infection (DPI), the mice were administered intraperitoneally with human plasma. A single dose of 0.2 mL plasma was given to those in group C. The mice from group D were administered five doses of 0.2 mL plasma with a 24 hours interval between the doses. Group B showed high increasing parasitemia that led to their death within 5 DPI. Both treatments eliminated parasites from the blood and increased the longevity of animals. An efficacy of 50 (group C) and 80% (group D) of human plasma trypanocidal activity was found using PCR. This therapeutic success was likely achieved in the group D due to their higher levels of APOL1 compared with group C.

scholarly journals Zeolite as a Bone Bio-Modifier Carrier: An In-Vitro Study

2019 ◽  
Vol 9 (1) ◽  
pp. 126
Author(s):  
Mobina Mousavi ◽  
Azadeh Esmaeil Nejad ◽  
Erfan Shamsoddin ◽  
Mohammad Mehdi Golabgiran ◽  
Behzad Houshmand
Keyword(s):  
In Vitro Study ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Alizarin Red ◽  
Group A ◽  
Group B ◽  
And Function ◽  
Group D

Background: Zeolite is a microporous aluminosilicate compound which has been successfully used in tissue engineering. The effects of Zeolite on the morphology and functions of pre-osteoblastic MG-63 cells as new bone enhancer material is still unclear. Methods: In this vitro experimental study, MTT and Alizarin red staining test were performed on six groups of MG-63 cells which differed in Zeolite (Z) concentration and the presence or absence of Alloplast extract (A). Group A: 0.1μg/mL Z+A, Group B: 0.1μg/mL Z without A, Group C: 0.2μg/mL Z+A, Group D: 0.2μg/mL Z without A, Group E: 0.3μg/mL Z+A, Group F: 0.3μg/mL Z without A. There were also three control groups as positive control, negative control, and Alloplast control based on each related test. The data were analyzed by SPSS 20 via one-way ANOVA and Welch test. (P<0.05). Results: At 24 hours, results showed that solutions with 0.1μg/mL, 0.2μg/mL, and 0.3μg/mL Zeolite with or without Alloplast had significantly higher proliferation rates than positive control (distilled water) groups without Alloplast (p<0.001). At 72hours time point, the results showed significantly higher proliferation rates in the solutions with 0.1μg/mL, 0.2μg/mL, and 0.3μg/mL Zeolite with or without Alloplast compared to the positive control group without Alloplast (p<0.001). Conclusions: Zeolite can increase proliferation of MG-63 cells without presence of Alloplast; It seems that combination of Zeolite with Alloplast maybe enhancing proliferation and function of MG-63 cells.

Trypanocidal activity of free and nanoencapsulated curcumin on Trypanosoma evansi

Parasitology ◽  
2014 ◽  
Vol 142 (3) ◽  
pp. 439-448 ◽  
Author(s):  
L. T. GRESSLER ◽  
C. B. OLIVEIRA ◽  
K. CORADINI ◽  
L. DALLA ROSA ◽  
T. H. GRANDO ◽  
...  
Keyword(s):  
Lethal Effect ◽  
Positive Control ◽  
Trypanosoma Evansi ◽  
Control Group ◽  
Trypanocidal Activity ◽  
Male Wistar Rats ◽  
Protein Peroxidation ◽  
Liver And Kidney

SUMMARYThis study aimed to evaluate in vitro and in vivo trypanocidal activity of free and nanoencapsulated curcumin against Trypanosoma evansi. In vitro efficacy of free curcumin (CURC) and curcumin-loaded in lipid-core nanocapsules (C-LNCs) was evaluated to verify their lethal effect on T. evansi. To perform the in vivo tests, T. evansi-infected animals were treated with CURC (10 and 100 mg kg−1, intraperitoneally [i.p.]) and C-LNCs (10 mg kg−1, i.p.) during 6 days, with the results showing that these treatments significantly attenuated the parasitaemia. Infected untreated rats showed protein peroxidation and an increase of nitrites/nitrates, whereas animals treated with curcumin showed a reduction on these variables. As a result, the activity of antioxidant enzymes (superoxide dismutase and catalase) differs between groups (P<0·05). Infected animals and treated with CURC exhibited a reduction in the levels of alanine aminotransferase and creatinine, when compared with the positive control group. The use of curcumin in vitro resulted in a better parasitaemia control, an antioxidant activity and a protective effect on liver and kidney functions of T. evansi-infected adult male Wistar rats.

scholarly journals Photopolymerizable Injectable Cartilage Mimetic Hydrogel for the Treatment of Focal Chondral Lesions: A Proof of Concept Study in a Rabbit Animal Model

2018 ◽  
Vol 47 (1) ◽  
pp. 212-221 ◽  
Author(s):  
Cecilia Pascual-Garrido ◽  
Elizabeth A. Aisenbrey ◽  
Francisco Rodriguez-Fontan ◽  
Karin A. Payne ◽  
Stephanie J. Bryant ◽  
...  
Keyword(s):  
Animal Model ◽  
Chondrogenic Differentiation ◽  
Osteochondral Lesions ◽  
Chondral Defect ◽  
Group A ◽  
Group B ◽  
Safranin O

Background: In this study, we investigate the in vitro and in vivo chondrogenic capacity of a novel photopolymerizable cartilage mimetic hydrogel, enhanced with extracellular matrix analogs, for cartilage regeneration. Purpose: To (1) determine whether mesenchymal stem cells (MSCs) embedded in a novel cartilage mimetic hydrogel support in vitro chondrogenesis, (2) demonstrate that the proposed hydrogel can be delivered in situ in a critical chondral defect in a rabbit model, and (3) determine whether the hydrogel with or without MSCs supports in vivo chondrogenesis in a critical chondral defect. Study Design: Controlled laboratory study. Methods: Rabbit bone marrow–derived MSCs were isolated, expanded, encapsulated in the hydrogel, and cultured in chondrogenic differentiation medium for 9 weeks. Compressive modulus was evaluated at day 1 and at weeks 3, 6, and 9. Chondrogenic differentiation was investigated via quantitative polymerase reaction, safranin-O staining, and immunofluorescence. In vivo, a 3 mm–wide × 2-mm-deep chondral defect was created bilaterally on the knee trochlea of 10 rabbits. Each animal had 1 defect randomly assigned to be treated with hydrogel with or without MSCs, and the contralateral knee was left untreated. Hence, each rabbit served as its own matched control. Three groups were established: group A, hydrogel (n = 5); group B, hydrogel with MSCs (n = 5); and group C, control (n = 10). Repair tissue was evaluated at 6 months after intervention. Results: In vitro, chondrogenesis and the degradable behavior of the hydrogel by MSCs were confirmed. In vivo, the hydrogel could be delivered intraoperatively in a sterile manner. Overall, the hydrogel group had the highest scores on the modified O’Driscoll scoring system (group A, 17.4 ± 4.7; group B, 13 ± 3; group C, 16.7 ± 2.9) ( P = .11) and showed higher safranin-O staining (group A, 49.4% ± 20%; group B, 25.8% ± 16.4%; group C, 36.9% ± 25.2%) ( P = .27), although significance was not detected for either parameter. Conclusion: This study provides the first evidence of the ability to photopolymerize this novel hydrogel in situ and assess its ability to provide chondrogenic cues for cartilage repair in a small animal model. In vitro chondrogenesis was evident when MSCs were encapsulated in the hydrogel. Clinical Relevance: Cartilage mimetic hydrogel may offer a tissue engineering approach for the treatment of osteochondral lesions.

scholarly journals Essential oils as tick repellents on clothing

2019 ◽  
Vol 79 (2) ◽  
pp. 209-219 ◽  
Author(s):  
Oliver Soutar ◽  
Freya Cohen ◽  
Richard Wall
Keyword(s):  
Essential Oils ◽  
No Reduction ◽  
Positive Control ◽  
Negative Control ◽  
Tick Abundance ◽  
Spearmint Oil ◽  
Oregano Oil ◽  
Negative Controls ◽  
Tick Repellents

Abstract Essential oils show promise as natural alternatives to synthetic tick repellents, but few studies have investigated their repellent efficacy in vivo or under field conditions. Here, blanket-drags and standardised walks were employed to evaluate tick acquisition by 1 m2 cotton blankets or cotton trousers, respectively, in woodland edge habitats of known high tick abundance. Blankets and trousers had been treated with one of 5% oregano, rosemary, spearmint or thyme oils, 20% DEET (N,N-diethyl-3-methylbenzamide) (positive control) or ethanol excipient-only (negative control). The number of ticks present on the blankets or trousers differed significantly between treatments: spearmint oil treatments resulted in significantly fewer ticks than the negative controls for both blankets and trousers and significantly fewer ticks were present on the oregano oil treated blankets. For ticks that did attach to the trousers, the rate of drop off within 3 min was significantly higher for trousers treated with spearmint oil or thyme oil than ethanol, oregano oil and rosemary oil. No reduction in repellence was detected over a 24 h period between treatment and testing. The results suggest that 5% oregano and spearmint oils exhibit potential as natural clothing repellents, with an effective equivalence to 20% DEET.

Effects of Removal of the Statoacoustic Ganglion Complex upon the Growing Otocyst

1976 ◽  
Vol 85 (6_suppl) ◽  
pp. 2-32 ◽  
Author(s):  
Thomas R. Van De Water
Keyword(s):  
Organ Culture ◽  
Inner Ear ◽  
Nerve Fibers ◽  
Trophic Effect ◽  
Group A ◽  
Sensory Structures ◽  
Statoacoustic Ganglion ◽  
Group B

An experiment was designed to answer the question as to whether or not the neural elements of the statoacoustic ganglion complex have a trophic effect upon the histodifferentiation of the sensory structures of the embryonic mouse inner ear anlage as it develops in vitro. The embryonic inner ear anlage with associated otic mesenchyme and statoacoustic ganglion complex was excised from 11, 12, and 13-day CBA/C57 mouse embryos. The inner ear explants of each gestational age group were further divided into two groups: the first group “A” (with) statoacoustic ganglion was explanted to the organ culture system without further surgical intervention; the second group “B” (without) statoacoustic ganglion underwent further surgical manipulation during which their statoacoustic ganglion complexes were dissected away prior to explantation to in vitro. The explanted embryonic inner ears were allowed to develop in organ culture until the equivalent of gestation day 21 in vivo was reached for each group; then all cultures were fixed and histologically processed and stained by a nerve fiber stain, in combination with a stain for glucoprotein membranes. Each specimen was code labeled and scored for histodifferentiation of sensory structures. Light microscopic observations confirmed that in group “A” cultures, statoacoustic ganglion neurons and their nerve fibers were present in association with the developed sensory structures; neither ganglion cell neurons nor their nerve fibers were found to be present in the sensory structures that developed in the group “B” organ culture specimens. Quantification revealed no consistent trend of greater occurrence of any sensory structure in the groups of explants analyzed. The presence of such a trend would have signified the probable existence of a trophic effect of the statoacoustic ganglion neural elements upon development of inner ear sensory structures in the group “A” explants of the 11, 12, and 13-day embryo inner ear organ culture specimens when compared to the aganglionic group “B” cultures. Microscopic comparison of the sensory structures and their sensory hair cells that developed in the organ cultures revealed no differences in the quality of the histodifferentiation of either group “A” or group “B” explants. A base to apex pattern of histodifferentiation of the organ of Corti sensory structures, which has been described to occur in vivo, was noted to occur in the in vitro developed cochlear ducts of all of the explanted inner ears without respect to whether neural elements were present (“A”) or absent (“B”) during development. It was concluded from the quantification of histodifferentiation data and the above observation on the pattern of differentiation of Corti's organ that no trophic effect of neural elements of the statoacoustic ganglion complex influencing the histodifferentiation of sensory structures of 11, 12, and 13-gestation day mouse embryo inner ear explants as they differentiate in vitro could be demonstrated.

scholarly journals Cleaved CD31 as a target for in vivo molecular imaging of inflammation

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Jonathan Vigne ◽  
Sylvie Bay ◽  
Rachida Aid-Launais ◽  
Guillaume Pariscoat ◽  
Guillaume Rucher ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Molecular Form ◽  
Positive Control ◽  
Negative Control ◽  
Stability Study ◽  
Biodistribution Studies ◽  
Wide Range ◽  
Significant Difference

AbstractThere is a need for new targets to specifically localize inflammatory foci, usable in a wide range of organs. Here, we hypothesized that the cleaved molecular form of CD31 is a suitable target for molecular imaging of inflammation. We evaluated a bioconjugate of D-P8RI, a synthetic peptide that binds all cells with cleaved CD31, in an experimental rat model of sterile acute inflammation. Male Wistar rats were injected with turpentine oil into the gastrocnemius muscle two days before 99mTc-HYNIC-D-P8RI (or its analogue with L-Proline) SPECT/CT or [18F]FDG PET/MRI. Biodistribution, stability study, histology, imaging and autoradiography of 99mTc-HYNIC-D-P8RI were further performed. Biodistribution studies revealed rapid elimination of 99mTc-HYNIC-D-P8RI through renal excretion with almost no uptake from most organs and excellent in vitro and in vivo stability were observed. SPECT/CT imaging showed a significant higher 99mTc-HYNIC-D-P8RI uptake compared with its analogue with L-Proline (negative control) and no significant difference compared with [18F]FDG (positive control). Moreover, autoradiography and histology revealed a co-localization between 99mTc-HYNIC-D-P8RI uptake and inflammatory cell infiltration. 99mTc-HYNIC-D-P8RI constitutes a new tool for the detection and localization of inflammatory sites. Our work suggests that targeting cleaved CD31 is an attractive strategy for the specific in vivo imaging of inflammatory processes.

scholarly journals Brazilian Red Propolis Is as Effective as Amoxicillin in Controlling Red-Complex of Multispecies Subgingival Mature Biofilm In Vitro

Antibiotics ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 432
Author(s):  
Kadmo Azevedo de Figueiredo ◽  
Helio Doyle Pereira da Silva ◽  
Stela Lima Farias Miranda ◽  
Francisco Jerfeson dos Santos Gonçalves ◽  
Arlene Pereira de Sousa ◽  
...  
Keyword(s):  
Metabolic Activity ◽  
Positive Control ◽  
Negative Control ◽  
In Vivo Studies ◽  
Complex Species ◽  
Hybridization Data ◽  
Cell Counts ◽  
Actinomyces Species

This study investigated the effects of Brazilian Red Propolis (BRP) extract on seven-day-old multispecies subgingival biofilms. Mixed biofilm cultures containing 31 species associated with periodontal health or disease were grown for six days on a Calgary device. Then, mature biofilms were treated for 24 h with BRP extract at different concentrations (200–1600 µg/mL), amoxicillin (AMOXI) at 54 µg/mL (positive control) or vehicle (negative control). Biofilm metabolic activity was determined by colorimetry, and bacterial counts/proportions were determined by DNA–DNA hybridization. Data were analyzed by Kruskal–Wallis and Dunn’s tests. Treatment with BRP at 1600, 800 and 400 μg/mL reduced biofilm metabolic activity by 56%, 56% and 57%, respectively, as compared to 65% reduction obtained with AMOXI. Mean total cell counts were significantly reduced in all test groups (~50–55%). Lower proportions of red, green and yellow complex species were observed upon treatment with BRP (400 µg/mL) and AMOXI, but only AMOXI reduced the proportions of Actinomyces species. In conclusion, BRP extract was as effective as AMOXI in killing seven-day-old multispecies biofilm pathogens and did not affect the levels of the host-compatible Actinomyces species. These data suggest that BRP may be an alternative to AMOXI as an adjunct in periodontal therapy. In vivo studies are needed to validate these results.

scholarly journals In vivo activities of ceftriaxone and vancomycin against Borrelia spp. in the mouse brain and other sites.

1996 ◽  
Vol 40 (11) ◽  
pp. 2632-2636 ◽  
Author(s):  
R J Kazragis ◽  
L L Dever ◽  
J H Jorgensen ◽  
A G Barbour
Keyword(s):  
Body Weight ◽  
Lyme Disease ◽  
Positive Control ◽  
Negative Control ◽  
Scid Mice ◽  
Immunodeficient Mice ◽  
Infected Adult ◽  
The Brain

Borrelia burgdorferi, the agent of Lyme disease, and B. turicatae, a neurotropic agent of relapsing fever, are susceptible to vancomycin in vitro, with an MIC of 0.5 microgram/ml. To determine the activity of vancomycin in vivo, particularly in the brain, we infected adult immunocompetent BALB/c and immunodeficient CB-17 scid mice with B. burgdorferi or B. turicatae. The mice were then treated with vancomycin, ceftriaxone as a positive control, or normal saline as a negative control. The effectiveness of treatment was assessed by cultures of blood and brain and other tissues. Ceftriaxone at a dose of 25 mg/kg of body weight administered every 12 h for 7 to 10 days eliminated cultivable B. burgdorferi or B. turicatae from all BALB/c or scid mice in the study. Vancomycin at 30 mg/kg administered every 12 h was effective in eliminating infection from immunodeficient mice if treatment was started within 3 days of the onset of infection. If treatment with vancomycin was delayed for 7 days or more, vancomycin failed to eradicate infection with B. burgdorferi or B. turicatae from immunodeficient mice. The failure of vancomycin in eradicating established infections in immunodeficient mice was associated with the persistence of viable spirochetes in the brain during antibiotic treatment.

scholarly journals Combination of garlic - shatterstone herb powder to control Streptococcus agalactiae infection in tilapia

2015 ◽  
Vol 14 (1) ◽  
pp. 79
Author(s):  
Ririn Nurul Fauziah ◽  
Dinamella Wahjuningrum ◽  
, Sukenda ◽  
, Ranta
Keyword(s):  
Growth Rate ◽  
Streptococcus Agalactiae ◽  
Allium Sativum ◽  
Positive Control ◽  
Quality Parameters ◽  
Negative Control ◽  
Fish Pathogen ◽  
Phyllanthus Niruri

<p class="NoParagraphStyle" align="center"><strong>ABSTRACT</strong><strong></strong></p><p class="NoParagraphStyle" align="center"><strong> </strong></p><p class="NoParagraphStyle">This study was aimed at determining potential of combination powder of garlic <em>Allium sativum</em>-shatterstone herb <em>Phyllanthus niruri</em> supplemented in feed against <em>S. agalactiae</em> infection in tilapia. Four concentrations of combination powder of <em>A. sativum</em>-<em>P. Niruri</em>; 20+5, 20+10, 20+15 and 20+20 ppt respectively were investigated for their ability to inhibit bacterial fish pathogen. Combination dose of 20+15 ppt produced the highest inhibitory zones in <em>in vitro</em> test. <em>In vivo</em> test consisted of three treatments with three replications, namely positive control (K+), negative control (K-) and the treatment of <em>A. sativum</em>-<em>P. niruri</em> suplemented in feed (BM).  The test perfomed on tilapia with weight of 10.33 ± 1.63 g and were reared at density of 10 ind/aquarium. The fish was fed for 14 days, then injected intraperitoneally with 0.1 mL <em>S. agalactiae</em> at concentration of 10<sup>5</sup> cfu/mL for positive control and BM groups. Survival, growth rate, feed response, hematological and water quality parameters were observed for 10 days. This study showed that the suplemented-feed-fish (BM) showed better growth rate, feed response, and survival (83.3%) than positive control (36.7%) at P&lt;0.05. In addition, <em>A. sativum</em>-<em>P. niruri</em> suplemented in feed was also able to enhance the immune response by increasing phagocytic activity.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: <em>Streptococcus agalactiae</em>, phytopharmacy, <em>Allium sativum</em>-<em>Phyllanthus niruri</em>, tilapia</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle" align="center"><strong>ABSTRAK</strong></p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Penelitian ini bertujuan untuk menganalisis potensi campuran tepung bawang putih <em>Allium sativum</em>-meniran <em>Phyllanthus niruri </em>dalam pakan terhadap pencegahan infeksi bakteri <em>S. agalactiae</em> pada ikan nila. Empat konsentrasi campuran tepung bawang putih-meniran yaitu 20+5 ppt, 20+10 ppt, 20+15 ppt dan 20+20 ppt masing-masing diuji kemampuannya dalam  menghambat bakteri patogen pada ikan. Campuran dosis 20+15 ppt menghasilkan zona hambat terbaik dalam uji <em>in vitro</em>. Uji <em>in vivo</em> terdiri atas tiga perlakuan dengan tiga ulangan yaitu kontrol positif, kontrol negatif, dan perlakuan pakan yang mengandung bawang putih-meniran (BM). Uji ini dilakukan pada ikan nila berbobot 10,33±1,63 g yang dipelihara di akuarium dengan kepadatan 10 ekor/akuarium. Ikan diberi pakan perlakuan selama 14 hari kemudian diinjeksi secara intraperitoneal dengan bakteri <em>S. agalactiae</em> sebanyak 0,1 mL dengan kepadatan 10<sup>5 </sup>cfu/mL pada perlakuan kontrol positif dan perlakuan BM. Parameter kelangsungan hidup, laju pertumbuhan, respons pakan, parameter hematologi, dan kualitas air diamati selama sepuluh hari. Hasil dari penelitian ini menunjukkan bahwa pemberian BM dalam pakan memberikan laju pertumbuhan, respons pakan, dan sintasan (83,3%) yang lebih baik daripada kontrol positif (36,7%) pada P&lt;0,05. Pakan yang mengandung campuran bawang putih-meniran ini juga mampu meningkatkan respons imun dengan adanya peningkatan aktivitas fagositosis.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Kata kunci: <em>Streptococcus agalactiae</em>, fitofarmaka, <em>Allium sativum</em>-<em>Phyllanthus niruri</em>, ikan nila</p><p> </p>

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