Beehive Products Research Articles

Trace element levels in an area impacted by old mining operations and their relationship with beehive products

The environmental status of an area impacted by Roman mining activities was assessed in order to establish the current risks posed by such old mine emplacements. For this purpose, soil samples were collected throughout the mining area and analysed for their total, mobile and mobilizable trace element (As, Cd, Mo, Sb and Zn) contents. Additionally, beehive products (honey and pollen) were also sampled and evaluated for their use as environmental indicators of the area. The results obtained were compared with those from a control non-polluted area. The mine soils presented slightly increased levels of Cd and Sb (about 2- to -3-fold their normal soil concentrations), whereas the enrichment of As reached considerable levels, with concentrations almost ten-fold of those considered the threshold for causing toxicity. Leachable As contents exhibited very high values (1.2–21.9mgkg−1), indicating the need for risk attenuation measures. All trace elements were mainly partitioned in the soil residual fraction, especially Mo (76–99%) and Sb (61–91%). Significant partitioning levels were also found in the reducible fraction of As (up to 35%) and Cd (up to 38%), and in the oxidizable fraction of Mo (up to 23%). The reducible pool of As was particularly relevant due to the eventual mobilization of this element under reducing conditions. Among the beehive products tested, honey proved not to be useful as an environmental indicator, whereas pollen showed great potential as an indicator when the contamination levels were moderate to high.

Natural strategies for the control of Paenibacillus larvae, the causative agent of American foulbrood in honey bees: a review

American foulbrood (AFB) is a severe bacterial disease that affects larvae of honey bees (Apis mellifera). The causative agent of AFB is the spore-forming bacteria Paenibacillus larvae. The use of antibiotics for the control of AFB has led to the appearance of resistant bacterial strains and residues in beehive products. Nowadays, antibiotics are legally banned in several countries, and the affected colonies have to be destroyed by burning the hives. Therefore, the development of alternative methods for the control and prevention of AFB is necessary. In this context, different natural strategies based on the application of essential oils, plant extracts, propolis, royal jelly, nonconventional natural molecules, bacteria, and bacteriocines, have been studied in vitro and in vivo for the prevention and control of P. larvae. The experimental data achieved from these studies are reviewed and discussed in the present review, which intend to be a starting point for future research in the field.

Indian Propolis Loaded Folic Acid Conjugated PLGA Nanoparticles: Formulation Development, Characterization, In Vitro and In Vivo Anticancer Study

Propolis, a natural bee hive product is well proven for its anticancer potential due its various polyphenols and flavonoids constituents. Anticancer efficacy of propolis is limited due to its poor water solubility and bioavailability. The present study is investigated for design and development of ethanolic extract of Indian propolis (EEIP) loaded folic acid conjugated Poly (D,L-lactide-co-glycolide) nanoparticles (denoted as ELFPN) were investigated to achieve improved solubility, sustained drug release and to study synergized anticancer efficacy. Formulation development, characterization and optimization were carried out by design of experiment approach. . In vitro and in vivo cytotoxicity study was carried out for optimized formulation. Developed ELFPN showed the particle size and encapsulation efficiency 178 ± 5 - 205 ± 5 nm and 73.16 ± 1.89 - 76.37 ± 1.89 respectively. Optimized formulation showed sustained drug release over a period of 48 h with no sign of blood toxicity. Moreover, concentration of the drug needed for growth inhibition of 50 % of cells in a designed time period (GI50) was decreased by 43.34 %, for ELFPN as compared to EEIP in human breast cancer MCF-7 cells indicating targeting with synergistic effect of ELFPN. An improved anticancer effect was reflected in in-vivo Daltons Ascites Lymphoma model by reducing tumor cells count. The developed ELFPN showed improved in vitro cytotoxic effect, in-vivo anti-cancer activity with desirable characteristics for nanoparticle formulation thus can be useful for biomedical applications.

Harmonização de metodologias de análise da própolis

Propolis (bee glue) is a resinous bee hive product used by man since ancient times due to its pharmaceutical properties. The variability of its chemical composition, linked with the floral origin, requires the analysis of several quality parameters. Nevertheless, at the European level, in addition to the inexistence of legal requirements for quality control, there is also a lack of analytical procedures, despite the presence in the literature of several methodologies, which are often not comparable. The International Honey Commission, an worldwide network aiming to implement and develop new analytical methods for the control bee products and the establishment of quality standards, defined as a priority, within the propolis working group, the harmonization of analytical procedures for propolis quality evaluation, particularly in respect to ash, wax (using two alternative methods), balsamic content after extraction with ultrasonic or mechanical agitation as well as the analysis of its phenolic composition through the assessment of total phenolics, flavones/flavonols and flavanones/di-hydroflavonols. This collaborative study was engaged by 15 entities, including 14 laboratories and an international company working with propolis. According to the results obtained in our laboratory it is possible to draw a first evaluation on the applied methods, in particular the simplicity on the protocol implementation, with exception for the wax content, which proved to be laborious and non-adapted for routine analysis. Nevertheless, the wax values from both methodologies seems to agree, independently of its level in the samples. For the extraction procedure (balsamic content), the samples showed values between 43% and 70%, independent of the use of mechanical stirring or sonication, which shows the advantage of this second method, which enables a significant reduction in labor time, regardless of the absolute value. Overall, the results found for the tested protocols, for the different parameters, revealed a constant performance, besides de natural oscillation associated with the floral origin.

Gc-ms analysis of phytocomponents in the methanolic extract of propolis of stingless bee

Propolis is a resinous Bee hive product and has been long used in folk medicine of different nations as early as 3000 BC. Stingless bees are indigenous pollinators of treasured herbal plants. Propolis of stingless bees is a natural, strongly adhesive mixture made of bee secretions and plant resins. Stingless bee Propolis exhibit a wide spectrum of pharmacological activities that vary depending on the geographical location, climate, generic type of bees and availability of botanical sources for the bees to forage. The present study was carried out to identify the phytocomponents present in the methanolic extract of the propolis of stingless bees reared and collected from Devanahalli, Bangalore, by GC-MS analysis. From the GC-MS results five compounds were identified as major constituents, they are Ethylhexanol, 1,1 dimethylethyl, , 4,6 dimethyl, , Diethyl phthalate, 1,2 benezenedicarboxylic acid. These different active phytochemicals have been found to possess a wide range of Pharmacological activities, which may help in the protection against incurable diseases.

Propolis induce cytotoxicity on cancerous hepatocytes isolated from rat model of hepatocellular carcinoma: Involvement of ROS-mediated mitochondrial targeting

Hepatocellular carcinoma (HCC) is the third cause of mortality due to cancer throughout the world. Propolis is a beehive product with a specific chemical composition, widely used in traditional medicine because of its several pharmacological activities. Propolis has attracted much attention in recent years as a useful substance having application in medicine and cosmetics. Many studies have shown that propolis contain anti-cancer compounds. The present research was designed to evaluate the selective cytotoxicity of ethanolic extracts of propolis (EEP) sample purchased from Kurdistan province, Iran on hepatocytes and mitochondria obtained from the liver of HCC induced rats. HCC was induced by diethylnitrosamine (DEN) and 2-acetylaminofluorene (2-AAF) method. Our results showed that EEP (75, 150 and 300μg/ml) selectively induced mitochondrial membrane potential (MMP)decrease, mitochondrial swelling and cytochrome c release by increasing reactive oxygen species (ROS) generation and also caspase-3 activation in hepatocytes and mitochondria isolated from liver of HCC induced but not untreated normal rats. Our results provided evidence for the hypothesis that Iranian propolis could show a promising selective toxicity on HCC rat hepatocytes through directly mitochondrial targeting and apoptosis induction.

Characterization of Lavandula spp. Honey Using Multivariate Techniques.

Traditionally, melissopalynological and physicochemical analyses have been the most used to determine the botanical origin of honey. However, when performed individually, these analyses may provide less unambiguous results, making it difficult to discriminate between mono and multifloral honeys. In this context, with the aim of better characterizing this beehive product, a selection of 112 Lavandula spp. monofloral honey samples from several regions were evaluated by association of multivariate statistical techniques with physicochemical, melissopalynological and phenolic compounds analysis. All honey samples fulfilled the quality standards recommended by international legislation, except regarding sucrose content and diastase activity. The content of sucrose and the percentage of Lavandula spp. pollen have a strong positive association. In fact, it was found that higher amounts of sucrose in honey are related with highest percentage of pollen of Lavandula spp.. The samples were very similar for most of the physicochemical parameters, except for proline, flavonoids and phenols (bioactive factors). Concerning the pollen spectrum, the variation of Lavandula spp. pollen percentage in honey had little contribution to the formation of samples groups. The formation of two groups regarding the physicochemical parameters suggests that the presence of other pollen types in small percentages influences the factor termed as “bioactive”, which has been linked to diverse beneficial health effects.

Essential minerals and inorganic contaminants (barium, cadmium, lithium, lead and vanadium) in dried bee pollen produced in Rio Grande do Sul State, Brazil

Like other beehive products, such as honey, royal jelly and propolis, bee pollen has attracted great interest because of the health benefits it can provide when consumed. Bee pollen has high contents of sugars and proteins and a low content of lipids, it is also a rich source of vitamins and other bioactive compounds, which makes it an attractive micronutrient supplement. However, few studies have investigated its composition. Therefore, the aim of this study was to characterize the essential minerals and inorganic contaminants present in bee pollen produced at apiaries in Rio Grande do Sul State, Brazil. Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) revealed the presence of 8 essential minerals (calcium, iron, copper, chromium, manganese, molybdenum, phosphorus and zinc) in the 5 analyzed samples; 6 of them were in sufficiently high amounts to meet dietary requirements. Of the 5 inorganic contaminants assessed (barium, cadmium, lithium, lead and vanadium), only cadmium was present at levels over the International Honey Commission’s standards. All bee pollen samples showed a high content of the 8 essential minerals. Contamination usually results from the use of pesticides, fertilizers and other chemicals in agriculture; thus, monitoring of its levels must be included in bee pollen analysis.

SKIN TEST REACTIVITY TO BEE HIVE PRODUCTS (HONEY BEES, HONEY, ROYAL JELLY AND POLLEN)

Bee hive products such as honey bees, honey, royal jelly and pollen are widely consumed as a health supplement. There has been several cases of allergic reactions to bee hive products reported worldwide. Consumption of bee hive products is common among Malaysians, but the degree of its sensitization is not known. The aim of this study is to determine the prevalence of skin test reactivity to Apis mellifera, domesticated locally and the bee hive products such as honey bees, honey, royal jelly and pollen among a group of volunteers. Four types of honey collected from different locations with different plant sources; one sample of imported honey from Australia, one sample of royal jelly, four different sources of bee pollen, and honey bee derivatives were used to prepare allergen extracts for skin testing. 2522 volunteers were skin tested to these allergen extracts. 271 (10.75%) of the subjects had a positive skin prick test result to at least one honey bee derivative or bee hive product allergen. Individuals with skin test positivity to honey bee are most likely to be sensitized to bee pollen followed by honey and royal jelly. Skin test reactivity to honey bee and bee hive products is prevailed to be high in Malaysia. This study has proven that bee hive products sensitization individuals are mostly to be also sensitized to honey bees. Therefore, is recommended these allergens to be included in the skin prick panel

Socio-economic profile and training needs of beekeepers in Samastipur district of Bihar

Bee benefit, the plants not only increasing their yield, but also improve crop quality since beekeeping does not compete for inputs with other farming or crop, it is an ideal programme for integration. Keeping the importance of beekeeping a study was undertaken to find out the socio-economic and psychological profile and training needs of beekeepers. The study highlighted that majority of the respondents belonged to middle to young age group, medium to high level of family education, socioeconomic status, innovativeness and risk bearing capacity. The results also showed that majority of respondents had low to medium level of extension contact and size of land holding, respectively. In respect to training needs of beekeepers in different areas, it was found highest in protection of bee pests, diseases and other hazards followed by the business of bees, beehive products and their extraction, processing and medicinal values, management of bee colonies during different seasons and essential operations.

Immunomodulatory activity of Apis mellifera propolis from the North of Argentina

Propolis is a bee hive product with complex chemical composition that was used as medicine from ancient times. The immunomodulatory activity of partially purified propolis extracts (PPPEs), galangin and pinocembrin was studied through their in vitro effect on neutrophil chemotactic and phagocytic activities. PPPEs (15 samples from each of 10 different beehives) were more effective (around 45% and 50% for a concentration of 40 μg of PPPEs/mL of active substance) as chemotactic agents than galangin and pinocembrin (around 20 and 25%, respectively) and showed higher neutrophil phagocitic activity (270 ± 10% for beehive 10) than galangin and pinocembrin (180 ± 10 and 135 ± 9%, respectively). The highest effect of PPPEs can be attributed to a synergistic effect among components of PPPEs. The results of this study indicated that PPPEs, galangin and pinocembrin stimulate neutrophil chemotactic activity at various concentrations suggesting their possible use for patients suffering neutrophil disfunction. The nitro blue tetrazolium results indicated that the extracts and isolated flavonoids have the capacity to scavenge active radicals suggesting that they would be valuable in the treatment of diseases associated with free radical damage.Consequently, the in vitro results suggest a potential value of these propolis extracts in the medical field.

Honey and beehive products in otorhinolaryngology: a narrative review

Honey and beehive products were rediscovered as an alternative treatment in wounds. The medicinal properties also raised interest of their use in Otorhinolaryngology. To give an overview of the effectiveness of beehive products in Otorhinolaryngology. Narrative. A literature search of the databases PubMed, EMBASE and Cochrane was performed from the last two decades till December 2014. The search terms 'honey', 'propolis' or 'royal jelly' were used. Articles, which evaluated the effectiveness of beehive products in Otorhinolaryngology, were included. The quality assessment of included studies was performed using the Cochrane Collaboration's risk of bias tool. A total of 36 studies were identified and evaluated. Eighteen studies investigated their effect in oral infections, seven in infection of the respiratory tract, six in rhino-sinusal diseases, four investigated the use in tonsillectomy and head and neck surgery and one study explored the preventive effect in otitis media. Honey can be considered as effective (additional) treatment in mucositis, childhood cough, persistent post-infectious cough and after tonsillectomy. Propolis may have a role in the treatment of (aphthous) stomatitis, mouth ulcer and prevention of acute otitis media. Royal jelly showed to reduce mucositis. In the presented studies, beehive products proved to be safe, with only minor adverse reactions. Studies showed to be diverse and had some methodological limitations.

Design of a food product composed of honey and propolis

Propolis is a natural bee hive product with potential functional activities. Nowadays, many propolis products are available on the market, including cosmetics and food commodities. However, it is still unknown if edible products with propolis are fully accepted by consumers and are thus commercially viable, due to the bitter and resinous flavor and taste of propolis. This work is focused on the design of a “honey with propolis” foodstuff that can be welcomed by consumers, and, if possible, provide added benefits compared to the honey itself. Propolis tinctures containing three different concentrations of ethanol and times of maceration were prepared, giving the best results with 90% ethanol and two days of maceration. Then, in order to study the maximum amount of propolis in honey that could be accepted by a representative group of consumers, a forced choice paired comparison preference test with 69 people was carried out, giving the best results with a honey product containing 0.3-0.5% propolis extract. Phenolic and flavonoid contents, as well as TEAC antioxidant and antimicrobial activities of propolis, honey, and the product “honey with propolis” were then analyzed. As expected, propolis showed the highest phenolics and flavonoids contents, as well as the highest antioxidant and antimicrobial activities. The product “honey with propolis” showed a significantly higher antimicrobial activity, however, when compared to honey alone. In conclusion, the commercialization of a food product of “honey and up to 0.5% propolis” is promising, interesting and viable, as such a product is greatly appreciated by consumers, as well as being potentially healthy.

Prochloraz and coumaphos induce different gene expression patterns in three developmental stages of the Carniolan honey bee (Apis mellifera carnica Pollmann)

The Carniolan honey bee, Apis mellifera carnica, is a Slovenian autochthonous subspecies of honey bee. In recent years, the country has recorded an annual loss of bee colonies through mortality of up to 35%. One possible reason for such high mortality could be the exposure of honey bees to xenobiotic residues that have been found in honey bee and beehive products. Acaricides are applied by beekeepers to control varroosis, while the most abundant common agricultural chemicals found in honey bee and beehive products are fungicides, which may enter the system when applied to nearby flowering crops and fruit plants. Acaricides and fungicides are not intrinsically highly toxic to bees but their action in combination might lead to higher honey bee sensitivity or mortality. In the present study we investigated the molecular immune response of honey bee workers at different developmental stages (prepupa, white-eyed pupa, adult) exposed to the acaricide coumaphos and the fungicide prochloraz individually and in combination. Expression of 17 immune-related genes was examined by quantitative RT-PCR. In treated prepupae downregulation of most immune-related genes was observed in all treatments, while in adults upregulation of most of the genes was recorded. Our study shows for the first time that negative impacts of prochloraz and a combination of coumaphos and prochloraz differ among the different developmental stages of honey bees. The main effect of the xenobiotic combination was found to be upregulation of the antimicrobial peptide genes abaecin and defensin-1 in adult honey bees. Changes in immune-related gene expression could result in depressed immunity of honey bees and their increased susceptibility to various pathogens.

Exploitation of Beehive Products, Plant Exudates and Tars in Corsica During the Early Iron Age

In the north‐western Mediterranean area, the first Iron Age is characterized by intense contacts and cultural interactions between populations. Archaeological remains such as ceramic vessels or metal and glass objects are usually good indicators of the nature and the intensity of these exchanges, but can also be used to determine the way in which these populations were living at their time. In contrast, organic substances, despite their importance in a wide variety of activities, are rarely investigated due to their low degree of preservation. The recent discovery of a series of amorphous organic residues with adhesive properties at the site of Cuciurpula provided a unique opportunity to address questions related to the types of natural substances exploited, their provenance, their uses and their informational input to intercultural relationships. Our results, based on GC and GC–MS analysis of organic residues preserved at the site of Cuciurpula, provide strong evidence for the most southern use of birch bark tar in Western Europe, and also for the simultaneous use of this substance with pine resin. Beeswax was also identified in some samples. The combined study of residue composition, aspect and location on ceramic sherds reveals a variety of uses, highlighting a complex technical system.

Propolis Ethanol Extract Stimulates Cytokine and Chemokine Production through NF-κB Activation in C2C12 Myoblasts.

Myoblast activation is a triggering event for muscle remodeling. We assessed the stimulatory effects of propolis, a beehive product, on myoblasts. After an 8 h treatment with 100 μg/mL of Brazilian propolis ethanol extract, expression of various chemokines, including CCL-2 and CCL-5, and cytokines, such as IL-6, increased. This propolis-induced cytokine production appears to depend on NF-κB activation, because the IKK inhibitor BMS-345541 repressed mRNA levels of CCL-2 by ~66%, CCL-5 by ~81%, and IL-6 by ~69% after propolis treatment. Supernatant from propolis-conditioned C2C12 cells upregulated RAW264 macrophage migration. The supernatant also stimulated RAW264 cells to produce angiogenic factors, including VEGF-A and MMP-12. Brazilian green propolis therefore causes myoblasts to secrete cytokines and chemokines, which might contribute to tissue remodeling of skeletal muscle.

Preparatory work for the pest categorisation of three bacteria listed in the annexes of the Council directive 2000/29/EC Erwinia amylovora Winslow et al. Xylophilus ampelinus (Panagopoulos) Willems et al. Clavibacter michiganensis subsp. insidiosus(McCulloch) Davis et al

Preparatory work for the pest categorisation of three bacteria listed in the annexes of the Council directive 2000/29/EC Erwinia amylovora Winslow et al. Xylophilus ampelinus (Panagopoulos) Willems et al. Clavibacter michiganensis subsp. insidiosus(McCulloch) Davis et al

Foreign matter contaminating ethanolic extract of propolis: a filth-test survey comparing products from small beekeeping farms and industrial producers

Propolis is a resinous material collected by honeybees from the exudates and buds of plants. It has been widely used as a remedy by humans since ancient times, as well as for dietary supplements and cosmetics. European legislation recently focused on the quality and hygiene standards of foods, including beehive products, and extensive efforts have been made to avoid the presence of chemical contaminants, whilst in contrast few studies have investigated the magnitude of contamination by physical ones. We conducted a filth-test survey to evaluate the contamination of ethanolic extract of propolis by foreign materials. We also compared the abundance of contaminants in propolis extracts currently marketed by small beekeepers and industrial producers. We found different foreign materials in the ethanol extract of propolis. Contaminants differed in abundance, with a higher number of carbon particles (small beekeepers: 2.70 ± 0.63; industrial producers: 1.25 ± 0.49; mean (n/30 ml) ± SE) and other inorganic fragments (small beekeepers: 3.50 ± 0.31; industrial producers: 3.88 ± 1.11) than arthropod fragments (small beekeepers: 0.30 ± 0.21; industrial producers: 0.38 ± 0.26) and mammal hairs (small beekeepers: 0.10 ± 0.10; industrial producers: 0.38 ± 0.26). No differences in the abundance of foreign matter between propolis from small beekeepers and industrial producers were found, allowing us to point out an increased awareness by small producers of issues inherent in hygiene management. Contamination of propolis extracts by animal body parts, such as insect fragments, mites and rodent hairs, indicates poor management of hygiene in the production process and low effectiveness of the filtration phase. Animal-borne contaminants can act as pathogen vectors as well as introducing dangerous allergens when ingested or applied to human skin. The filth-test applied to ethanolic propolis extract quality control can be considered a promising tool, also for small beekeeper activities, since it is cheap and allows the quick interpretation of results.

The immunomodulatory effect of propolis on receptors expression, cytokine production and fungicidal activity of human monocytes.

Propolis is a beehive product and its immunomodulatory action has been documented; however, little is known concerning its mechanisms of action on human cells. Propolis influence on the initial events of the immune response was assessed, evaluating cell markers, cytokine production and the fungicidal activity of human monocytes. Toll-like receptor (TLR)-2, TLR-4, human leukocyte antigen-DR and cluster of differentiation (CD)80 expression by human monocytes was assessed using a FACSCalibur flow cytometer, cytokine production (tumour necrosis factor (TNF)-α and interleukin (IL)-10) was determined by ELISA and the candidacidal activity was investigated after monocytes incubation with propolis and challenged with Candida albicans. The role of TLR-2 and TLR-4 on propolis action was assessed as well. Propolis upregulated TLR-4 and CD80 expression and affected TNF-α and IL-10 production, depending on concentration. Propolis also increased the fungicidal activity of monocytes. Cytokine production was decreased by blocking TLR-4, whereas the fungicidal activity was affected by blocking TLR-2. Propolis exerted an immunomodulatory action on cell receptors, cytokine production and fungicidal activity of human monocytes without affecting cell viability and depending on concentration. TLR-2 and TLR-4 may be involved in its mechanism of action.

Propolis extracts가 양식넙치의 면역활성능에 미치는 영향

Propolis extracts는 다양한 성분으로 구성되어 있는 수지상의 혼합물로서 여러 약리작용을 보여주기 때문에 민간요법으로 널리 이용되어왔다. 특히 propolis extracts는 독성이 거의 없다는 장점 때문에 면역증강제재로서 활용가능성이 높게 평가되고 있다. 이에 본 실험에서는 대상으로 in vitro 및 in vivo상에서 propolis extracts가 양식넙치 면역반응에 미치는 영향을 조사하였다. In vitro상에서 propolis extracts가 PBL의 탐식능 및 NBT assay활성능에 미치는 영향을 조사 한 바 유효농도는 <TEX>$50{\sim}150{\mu}g/ml$</TEX> 이었으며, 가장 활성이 높은 농도는 <TEX>$100{\mu}g/ml$</TEX> 이었다. In vivo상에서 양식넙치에 propolis extracts를 단독 또는 S. iniae 불활화 균액과 혼합하여 어체에 투여 후에 면역반응에 미치는 영향을 조사하였다. Haematocrit치는 propolis extracts단독 및 vaccine과 혼합 처리 할때 유의적 차이를 보이지 않았다. 그렇지만, 혈청 lysozyme과 신장대식세포의 탐식능, 활성산소 환원능 및 항체가는 propolis extracts+vaccine 투여구에서 유의적으로 증가하였다. 이러한 결과들에 의하여 propolis extracts는 양식넙치에서 특이적 및 비특이적 면역증강작용이 우수하므로 향후 양식어류에 있어서 비특이적 면역증강 물질 및 adjuvant로 활용가능성이 기대되어진다. Propolis is a beehive product with a very complex chemical composition, widely used in folk medicine because of its several therapeutic activities. This study was conducted to measure the efficacy of propolis on non-specific defense reactions, specific immune response, and protection levels against pathogen challenge with Streptococcus iniae. in vitro and in vivo. In vitro, the phagocytic activity and NBT assay of peripheral blood leukocytes (PBL) were evaluated in a various propolis extractsconcentrations (0, 10, 50, 100, 150, 250 and <TEX>$500{\mu}g/ml$</TEX>). The optimal concentration showing activation of propolis extracts was determined to <TEX>$100{\mu}g/ml$</TEX>. In vivo, they were divided into four groups (PBS, propoli extractss, vaccine, propolis extracts + vaccine) in vivo. Fish were received i.p. injection of either PBS or propolis extracts, and in the presence or absence of formalin inactivated S. iniae (<TEX>$1{\times}10^8$</TEX> CFU/fish), respectively. The level of haematocrit is not affected among experimental groups. The phagocytic activity and the NBT reduction activities of head kidney phagocyte were markedly (p<0.05)augmented in the propolis extracts groups than in the PBS-control group, respectively. The level of serum lysozyme activity was significantly (p<0.05) increased in the propolis extracts treated groups than in the PBS-control group. The agglutinin titer was significantly (p<0.05) enhanced in the vaccine+propolis extracts group than in the vaccine group, but there was no difference between PBS-control and propolis treated group. The results of the present study suggest that propolis extracts seems to be a promising compounds of non-specific immune stimulator, also being able to use a good adjuvant.