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ANALYSIS of POLLEN LOADS COLLECTED by HONEY BEES (APIS MELLIFERA L.) from LAMBAYEQUE PROVINCE (PERU): BOTANICAL ORIGIN and PROTEIN CONTENT Diana M

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ANALYSIS of POLLEN LOADS COLLECTED by HONEY BEES (APIS MELLIFERA L.) from LAMBAYEQUE PROVINCE (PERU): BOTANICAL ORIGIN and PROTEIN CONTENT Diana M Journal of Global Biosciences ISSN 2320-1355 Vol. 3(1), 2014, pp. 285-298 http://mutagens.co.in ANALYSIS OF POLLEN LOADS COLLECTED BY HONEY BEES (APIS MELLIFERA L.) FROM LAMBAYEQUE PROVINCE (PERU): BOTANICAL ORIGIN AND PROTEIN CONTENT Diana M. Saavedra-Carhuatocto 1, Francisco Aguinaga-Castro 1† 2 , Consuelo Rojas-Idrogo 2 & 2 Guillermo E. Delgado-Paredes 1Facultad de Ingeniería Zootecnia, Universidad Nacional Pedro Ruiz Gallo, Ciudad Universitaria, Juan XXIII N o 391, Lambayeque, PERÚ. †In Memoriam 2Facultad de Ciencias Biológicas, Universidad Nacional Pedro Ruiz Gallo, Ciudad Universitaria, Juan XXIII N o 391, Lambayeque, PERÚ. Abstract In the localities of Illimo, Motupe and Olmos (Phytogeographical province of Lambayeque, Peru in the equatorial dry forest) the availability of flowering along with the botanical origin and protein content of pollen loads harvested by honey bees ( Apis mellifera ), was studied. Based on the melissopalynology 20 pollen load samples were studied and according to qualitative analysis, 14 samples were monofloral and 6 were multifloral. During one apiculture period (September to November) pollen loads were collected every fortnight using pollen traps. Twenty-three principal pollen types were identified belonging to species occurring in the study area. The families Fabaceae and Capparaceae had a significant importance amongst the samples, represented by five and three pollen types, respectively. Prosopis limensis and Acacia macracantha pollen types occurred most constantly among the samples. A total of 200 g of pollen loads was analyzed. Pollen types such as Prosopis limensis , Capparis scabridae , C. avicennifolia and Acacia macracantha are considered the indicators for this geographical origin. The studied pollen loads were also characterized by a high frequency of Spilanthes leiocarpa , Alternanthera peruviana , Baccharis glutinosa and Parkinsonia aculeata . P. limensis reached the largest amount of pollen grains with 55 812.5 grains/mm 3 (58.9% of frequency) in Olmos and was the dominant pollen ( ≥ 45%) in the same locality. The highest content of crude protein and amino acids, especially methionine and histidine, also correspond to pollen load samples of Olmos locality. The high representation of native flora distinguishes these pollen loads geographically . Key words: Honey bees, pollen loads, pollen morphology, protein content, seasonally dry forest . Date of Online: 28-02-2014 INTRODUCTION One of the most known and studied plant-insect relationship models is between bees and melliferous plants. Due to their harvesting habit, bees are considered to be the most important and efficient plant pollinating insects, and in natural conditions play and important role in maintaining biodiversity of plants [1]. Indeed bees harvest ten to 1000 times more pollen than other insects. Individual bees leave the hive 20 to 30 times a day and the hive’s bees may visit 20 to 30 million flowers during a season [2]. Most plants are reproductively dependent upon the activities of the pollinating insects for transportation of pollen from one plant to another, for this reason, this symbiotic relationship ensures reproduction and genetic diversity for the plants, while the insects gain nectar and pollen as food from the plants [3]. In fact, pollen is a source of proteins, free amino acids, lipids, minerals and vitamins which are essential to growth and development of honey bees rather than energy production [4]. In particular, nitrogen is crucial for development of larvae and longevity of adults [5, 6]. It is important to detect which are the main pollen sources of a region and to determine their protein value while pollen is a major component of honey bees’ diet. Apies mellifera L. transports pollen to the hives in specialized Journal of Global Biosciences Vol. 3(1), 2014 pp. 285-298 ISSN 2320-1355 structures of its legs (corbicula), in which pollen, moistened with nectar and saliva, is packaged forming pasty “pellets”, called ‘pollen loads’ [7]. These loads are generally monoespecific due to the high grade of floral constancy of A. mellifera [8]. Melissopalynology, the microscopic analysis of pollen from honey sediment and pollen loads, was the first method used for determining the botanical and, in some cases, the geographical origin, therefore, more valuable information about the characterization of honeys and pollen loads can be obtained from melissopalynological studies [9]. This information is helpful for developing an apiary industry and commercial honey production [10]. In addition, apiculture is becoming an increasingly important socio-economic activity in several areas of Peru and particularly in the equatorial dry forest, a biome unique in the world stretching from the Santa Elena peninsula in Ecuador (0 o30’ South Latitude) to the middle basin of the Chicama river (La Libertad region) in the north of Peru (5 o00’ South Latitude); however, there are not studies on the potential nectar sources and identification of melliferous plants in this ecosystem , consequently, the level of effective use of the vegetation by bees is still to be determined. Characterization of monofloral origin has been recently initiated due to consumer demands. Tsigouri et al. (2004) [11] have emphasized that ‘‘the product name may be supplemented by information referring to floral or vegetable origin, if the product comes wholly or mainly from the indicated source and possesses the organoleptic, physicochemical and microscopic characteristics of the source’’ [12, 13]. Generally, blossom honey is considered to be from one source (monofloral) if the pollen frequency of that plant is more than 45% [14]. There has been a limited research on pollen analyses of Peru honeys and pollen loads and more are needed to determine the pollen and plant sources in different regions [15]; due to, for other countries are reported numerous studies. In Argentina, there are several studies on the botanical composition of pollen loads and pollen flora in the central region, where apiculture has a great developed [16, 17], and was recently reported the botanical composition and protein content of pollen collected by A. mellifera in the north-west of Santa Cruz (Argentinean Patagonia) [8]. In Spain an analysis was made of the protein content of pollen loads produced by the bees in a hive situated in Viana do Bolo (Ourense, north-west Spain), to establish whether or not the relative quantity of protein in the pollen of each plant species influences the preference made by the bee of the flowers that supply pollen to the hive [18]; likewise, was studied the seasonal variation in vegetation an pollen collected by honey bees in Thessaloniki region, Greece [19], and the seasonal variation of collected pollen loads of honey bees ( Apies mellifera anatoliaca ) in Bursa region, Turkey [20]. The aim of this study was to determine the diversity of plant species used as source of pollen by honey bees ( Apis mellifera ), to identify the morphological characteristics the pollen collected by honey bees and obtained from several beehives at Lambayeque province (Peru) as well as the protein content. MATERIALS AND METHODS Study area . The study area is located in the Lambayeque province (Lambayeque region), in the Peruvian north-coast, very next to the Pacific Ocean (79º27’ – 80º00’ West Longitude and 5º03’ – 6º09’ South Latitude) (Figure 1). Altitude is 500 m on average. Soil contents sand (10 – 40%), slime (30 – 60%) and clay (10 – 30%). The climate is characterized by a rainy season (December to March) with 100 – 500 mm of precipitation per year and a dry season between April and November; average relative humidity is 69%. Annual temperature is 19.5 and 31.2 oC. The natural vegetation is composed by the following zones: “El Chaparral” ( Cordia lutea , Capparis scabridae , Prosopis limensis and Capparis avicennifolia , as predominant species), “El Algarrobal” ( P. limensis , C. scabridae , C. lutea and Acacia macracantha , as predominant species), “El Zapotal” ( C. scabridae , as predominant species) and “El Gramadal” ( Spilanthes leiocarpa , Ambrosia peruviana and Lippia nodiflora , as predominant species) [21]. In general, the studies area corresponds to the equatorial dry forest [22] or seasonally dry forest, which has been deeply modified by a dense human population whose main activity is agriculture and firewood extraction. Agriculture activity is based on irrigation. Most of the cultivated area is planted with maize ( Zea mays ), common bean ( Phaseolus vulgaris ), cotton ( Gossypium hirsutum ), rice ( Oryza sativa ) and several fruits as Citrus sp. (lemons, oranges, limes and satsumas), passion flower ( Passiflora edulis ), mango ( Mangifera indica ) and avocado ( Persea americana ). Fooder crops, mainly alfalfa ( Medicago http://mutagens.co.in 286 Journal of Global Biosciences Vol. 3(1), 2014 pp. 285-298 ISSN 2320-1355 sativa ) and horticultural plants are cultivated in a lesser degree. Apiculture is complementary to agriculture as bees are used for the pollination of fruit crops. Reference slides of pollen plants . Plants were identified by Dr. Guillermo E. Delgado and stored in the herbarium HPR at the Universidad Nacional Pedro Ruiz Gallo (UNPRG), Lambayeque (Peru). Anthers were removed from flower buds of fresh plants, acetolyzed with glacial acetic acid, centrifuged, washed once with distilled water, centrifuged again, acetolyzed with anhydrous acetic acid and sulfuric acid [23, 24] mounted in glycerin jelly on glass slides and sealed with paraffin.
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