Printer Friendly

Thermal requirements and annual number of generations of Diachasmimorpha longicaudata (Hymenoptera: Braconidae) reared in the South American fruit fly and the Mediterranean fruit fly (Diptera: Tephritidae).

Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae), which originated in the Indo-Australian region, is a koinobiont endoparasitoid of various tephritid species. It has been introduced into various tropical and subtropical regions for control of Anastrepha species (Diptera: Tephritidae) and Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) (Montoya et al. 2000). This parasitoid is considered the most important biological control agent for inundative releases against diverse species of fruit flies in Latin American countries (Lopez et al. 2009).

In 1994 Embrapa (the Brazilian Corporation of Agricultural Research) introduced D. longicaudata into Brazil from the Florida Department of Agriculture and Consumer Services, Division of Plant Industry, Gainesville, Florida, USA, and released it in northeastern Brazil, where it is now established in the Reconcavo Baiano region and in the Subme dio Sao Franscisco region (Carvalho & Nascimento 2002). Its establishment has been demonstrated in the states of Minas Gerais (Alvarenga et al. 2005) and Rio de Janeiro (Leal et al. 2008). However, there are no records in regards to its release in Rio Grande do Sul.

Fruit flies (Diptera: Tephritidae) are first-order pests in fruit crops in Brazil (Nava & Botton 2010). In Rio Grande do Sul, C. capitata and Anastrepha fraterculus (Wiedemann) are important pests of exotic fruit crops such as apple and peach (Kovaleski & Ribeiro 2003). Anastrepha fraterculus also damages native fruit crops including several Myrtaceae (Gattelli et al. 2008).

Temperature is an important climatic variable that can directly influence the phenology and distribution of insects (Damos & Savopoulou-Soultani 2012). The optimal temperature for the development of a pest species may not be the same as that of its natural enemy (Horn 1998). Determination of the most appropriate temperature for the development of a parasitoid is important for its laboratory production and for planning field releases, and is a fundamental requirement of any program of biological control of pests. Meirelles et al. (2013) determined the development of D. longicaudata on C. capitata and A. fraterculus at a constant temperature (25 [+ or -] 1[degrees]C). This study sought to determine the thermal requirements of D. longicaudata reared in these 2 host species, to estimate the potential number of generations of D. longicaudata per year in 4 fruit-producing regions of the state of Rio Grande do Sul, and to reveal information about the development and possible establishment of this parasitoid in these regions.

Materials and Methods


All insects were reared at 25 [+ or -] 2[degrees]C, 65 [+ or -] 10% RH, and a photoperiod of 14:10 h L:D.

Fruit Flies. Adults of A. fraterculus and C. capitata were kept in separate cages (45 x 30 x 30 cm). Food-adapted from Salles (1995) including wheat germ, beer yeast, the hydrolyzed protein Biorigin[R], crystal sugar (at a ratio of 1:1:1:3), and water--was supplied ad libitum. Adults of A. fraterculus received oviposition substrates for 24 h (Salles 1992) composed of agar (5 g), blackberry juice (80 mL), and water (160 mL). The eggs were removed from the substrate and kept in distilled water for 48 h in a climate chamber (25 [+ or -] 2[degrees]C, 65 [+ or -] 10% RH; in the dark), and then transferred to a larval diet. A 250 mL orange plastic tube was used as a substrate for C. capitata oviposition, following the method of FAO/IAEA/USDA (2003). The eggs were removed daily from the substrate and held 24 h before being placed in a climate chamber on the larval diet. The larval diet for both fly species was composed of raw carrot (500 g), cooked carrot (500 g), crystal sugar (500 g), beer yeast (100 g), corn flour (600 g), nipagin (4.4 g), sodium benzoate (4.4 g), citric acid (14.4 g), and distilled water (400 mL) (Teran 1977). The diet was placed on trays, where the eggs were distributed. The trays with larval diet and eggs were wrapped in newspaper and held for 7 d. Then the trays were unwrapped, placed on sterilized sand, and held for an additional 10 d. During this time the larvae exited the diet and pupated in the sand. The sand was then sifted, and the pupae were placed in plastic boxes (15'15'5 cm) with sterilized sand until emergence.

Parasitoid. The first specimens of D. longicaudata were obtained from the laboratory of the Embrapa Mandioca e Fruticultura research institute (located in Cruz das Almas, Bahia, Brazil) in Oct 2008. The adults, approximately 50 couples, were kept in cages (20 x 15 x 15 cm) covered with voile. Water and food (a mixture of water, honey, sugar, ascorbic acid, and nipagin) were supplied ad libitum. Third instars of C. capitata were exposed for 1 h (40 to 50 larvae per parasitism unit) to adults of D. longicaudata. Parasitized larvae were placed in plastic boxes (15 x 15 x 5 cm) with sterilized sand until emergence.


Six sets of 50 pairs of D. longicaudata, between 3 and 10 d old, were kept in separate cages (20 x 15 x 15 cm), receiving food and water ad libitum. During several days, each set of D. longicaudata was exposed for 1 h to 40 to 50 third instars of A. fraterculus (~9 d old). About 18 sets of larvae were used. After exposure, the larvae were placed individually in glass tubes (25 mL) with sterilized sand and damp filter paper, and the tubes were sealed with plastic wrap and distributed evenly in chambers maintained at 15, 18, 21, 25, 28, and 31[degrees]C, at 60 [+ or -] 10% RH and a photoperiod of 14:10 h L:D. The insects that emerged at each temperature regime (between 109 and 200 individuals) were placed individually in plastic jars (140 mL) with food and water, and maintained at the same temperature regime until death. Adult emergence and mortality were recorded each morning. The same procedure was repeated exposing parasitoids to larvae of C. capitata.

The thermal requirements of the development periods (defined as the period of time from egg to adult emergence) of D. longicaudata at the 6 temperatures were estimated through the hyperbola method (Haddad et al. 1999), which calculates the lower development threshold (Dt), the thermal constant (K), and the optimum temperature range for development. The average development period and the longevity of the parasitoids reared in the same host at different temperatures were analyzed through the Kruskal-Wallis test (P [less than or equal to] 0.05) and Dunn's paired comparisons. The development period of parasitoids reared in the 2 different hosts at the same temperature regimes were compared by the Mann-Whitney test (P [less than or equal] 0.05) using the program BioEstat[R] 5.0 (Ayres et al. 2007). The possible number of generations of D. longicaudata per year was estimated for the counties of Bento Goncalves, Pelotas, Porto Alegre, and Vacaria, Rio Grande do Sul, based on the parasitoid's thermal requirements and the mean temperatures of these locations (Applied Meteorology Center of FEPAGRO and the 8th Meteorology District, Porto Alegre), according to Cividanes (2000).


No D. longicaudata completed development at either 15 or 31[degrees]C in A. fraterculus, or at 15[degrees]C in C. capitata (Table 1). The viability (completion of the egg-to-adult cycle) of D. longicaudata was greatest at 25[degrees]C when the parasitoid was reared in either host species, and this value was 61.3% in A. fraterculus and 58.3% in C. capitata. The mean development period of D. longicaudata reared in A. fraterculus was longest at 18[degrees]C and shortest at 28[degrees]C, and this period was inversely related to temperature and differed among all temperature regimes (H = 424.54; df = 3; P < 0.0001) (Table 1). When the host was C. capitata, the development period varied inversely with temperature among the temperatures of 18, 21, and 25[degrees]C (H = 603.04; df = 4; P < 0.0001) but was not significantly different between 28 and 31[degrees]C.

The development period of D. longicaudata was longer in A. fraterculus than in C. capitata at all temperatures. Females of D. longicaudata took more time to reach the adult phase than males at all temperatures, both in C. capitata (H = 634.96; df = 9; P < 0.0001) and in A. fraterculus (H = 439.62; df = 7; P < 0.0001).

The lower development threshold (Dt) of males and females of D. longicaudata in A. fraterculus (12.5[degrees]C) was higher than in C. capitata (7.83[degrees]C) (Table 2). Approximately 99% and 93% of the reduction of the development time at increasing temperatures of D. longicaudata reared in A. fraterculus and C. capitata, respectively, was explained by the increase in temperature (Table 2). The optimum temperature range for development of D. longicaudata was 16.37 to 22.48[degrees]C in A. fraterculus and 11.56 to 21.86[degrees]C in C. capitata.

An inverse relationship between the mean longevity of D. longicaudata and temperature was found. When D. longicaudata was reared in A. fraterculus, the longest development period was recorded at 18[degrees]C (52.7 d) and the shortest at 28[degrees]C (9.2 d). The longevities at 21[degrees]C (29.5 d) and 25[degrees]C (27.0 d) were statistically similar (Table 3). With C. capitata as its host, the parasitoid had statistically similar longevities at 18[degrees]C (53.3 d) and 21[degrees]C (39.4 d), intermediate longevity at 25[degrees]C (29.5 d), and the shortest longevities at 28[degrees]C (11.3 d) and 31[degrees]C (11.5 d). Individuals developing in C. capitata lived longer at 21[degrees]C (U = 4742.0; P < 0.0001) than those developing in A. fraterculus, i.e., 39.4 d vs. 29.5 d. There was no statistically significant difference between the longevities of D. longicaudata reared in the 2 hosts at 18, 25, or 28[degrees]C.

The estimated number of generations of D. longicaudata per year for the 4 counties (Table 4) when the host was A. fraterculus varied from 4.84 to 11.16, and the corresponding values for C. capitata as the host varied from 8.28 to 13.12. Thus D. longicaudata has the potential for more generations per year in C. capitata than in A. fraterculus.


The failure of D. longicaudata to produce adults at 15[degrees]C in both host species may be attributed to the existence of a dormancy phase stimulated by low temperature. Diapause in larvae of D. longicaudata collected in regions of Hawaii, USA, with winter temperatures varying between 2 and 10[degrees]C, was recorded by Clausen et al. (1965). However, the method utilized in our study did not permit us to observe this phenomenon. The failure of the parasitoid to develop at 31[degrees]C in A. fraterculus deserves further studies using individuals more experienced on that host. In C. capitata, at 28 and 31[degrees]C, emergence of D. longicaudata was similar to that recorded in Anastrepha suspensa (Loew) (Diptera: Tephritidae) by Ashley et al. (1976). The results obtained in our study may be associated with the fact that the parasitoid had been reared for many generations in C. capitata. The rearing of this parasitoid in Brazil has been conducted in C. capitata ever since its introduction in 1994. If D. longicaudata had been reared for many generations in A. fraterculus, it seems possible that its Dt value would be different from that reported here.

Vacaria has an annual average temperature (15.2[degrees]C) that is unfavorable to the parasitoid, regardless of the host, when compared with other regions; thus, fewer generations would be expected per year in this region. Among the 4 counties, Porto Alegre had the largest estimated number of generations per year. We expect that in warmer regions, or in periods with mean temperatures above 25[degrees]C, D. longicaudata would develop more rapidly, as the time necessary to double its population at 25[degrees]C is 3.92 and 4.73 d in A. fraterculus and C. capitata, respectively (Meirelles et al. 2013). Perhaps the success of D. longicaudata in regions of many countries where it has been introduced as a biological control agent can be explained in part by higher mean temperatures in these regions compared with the fruit-growing counties of Rio Grande do Sul.

During the winter in Rio Grande do Sul, during the months of Jun through Aug, temperatures may restrict or impair the establishment of D. longicaudata. Studies of population fluctuations of A. fraterculus in states of southern Brazil point to a reduction in populations during the months of Jun and Jul (Kovaleski et al. 2000), when the mean minimum temperatures are frequently below the development threshold for this parasitoid. The primary hosts of the South American fruit fly in Rio Grande do Sul are species of fruit trees in the Myrtaceae and Rosaceae (Zucchi 2000), which do not have mature fruits in Jun and Jul (Simao 1971; Donadio 2000; Manica 2002; Lira Junior et al. 2007). Between the maturation of fruit of feijoa (pineapple guava) (Acca sellowiana (O. Berg) Burret; Myrtales: Myrtaceae) in May, and the maturation of fruit of Brazilian cherry (Eugenia species; Myrtales: Myrtaceae) and loquat (Eriobotrya japonica [Thunb.] Lindl.; Rosales: Rosaceae) in Aug, there could be a significant scarcity of hosts for fruit flies and consequently for D. longicaudata. Individuals of D. longicaudata originating from C. capitata had a lower thermal threshold than those reared in A. fraterculus. The development rate of this parasitoid, reared in both hosts, increases with an increase in temperature. It is likely that in some regions of southern Brazil, D. longicaudata would have difficulty surviving through the winter, making necessary the releases of this parasitoid early in spring.


We thank the National Council for Scientific and Technological Development (CNPq) for fellowships awarded to the first and second authors (proc. n. 305956/2010-8) and for financial support (proc. n. 475287/2010-0). We would like to thank Antonio Nascimento at Embrapa Mandioca e Fruticultura Tropical for providing the first D. longicaudata individuals that we used to establish our laboratory colony.

References Cited

Alvarenga CD, Brito ES, Lopes EN, Silva MA, Alves DA, Matrangolo CAR, Zucchi RA. 2005. Introducao e recuperacao do parasitoide exotico Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae) em pomares comerciais de goiaba no norte de Minas Gerais. Neotropropical Entomology 34: 133-136.

Ashley TR, Greany PD, Chambers DL. 1976. Adult emergence in Biosteres (Opius) longicaudatus and Anastrepha suspensa in relation to the temperature and moisture concentration of the pupation medium. Florida Entomologist 59: 391396.

Ayres M, Ayres Jr M, Ayres DL, Dos Santos AS. 2007. BioEstat 5.0 aplicacoes estatisticas nas areas da ciencias biologicas e medicas. Sociedade Civil Mamiraua & CNPq, Belem, Brazil.

Carvalho R Da S, Nascimento AS. 2002. Criacao e utilizacao de Diachasmimorpha longicaudata para controle biologico de moscas-das-frutas, pp. 65-179 In Parra JRP, Botelho PSM, Correa-Ferreira BS, Bento JMS [eds.], Controle biologico no Brasil: Parasitoides e Predadores. Manole, Sao Paulo, Brazil.

Cividanes F. 2000. O uso dos graus-dia em entomologia: Com particular referencia ao controle de percevejos pragas da soja. Funep, Jaboticabal, Brazil.

Clausen CP, Clancy DW, Chock QC. 1965. Biological control of the oriental fruit fly (Dacus dorsalis Hendel) and other fruit flies in Hawaii. USDA, ARS, Technical Bulletin 1322: 1-102.

Damos P, Savopoulou-Soultani M. 2012. Temperature-driven models for insect development and vital thermal requirements. Psyche 2012: Article 123405.

Donadio LC. 2000. Novas variedades brasileiras de frutas. Sociedade Brasileira de Fruticultura Jaboticabal, Brazil.

FAO/IAEA/USDA. 2003. Manual for Product Quality Control and Shipping Procedures for Sterile Mass Reared Tephritid Fruit Flies, Version 5.0. International Atomic Energy Agency, Vienna, Austria.

Gattelli T, Silva FF Da, Meirelles RN, Redaelli LR, Dal Soglio FK. 2008. Moscas frugivoras associadas a mirtaceas e laranjeira 'Ceu' na regiao do Vale do Rio Cai, Rio Grande do Sul, Brasil. Ciencia Rural 38: 236-239.

Haddad ML, Parra JRP, Moraes RCB. 1999. Metodos para estimar os limites termicos inferior e superior de desenvolvimento de insetos. FEALQ Piracicaba, Brazil.

Horn DJ. 1998. Temperature synergism in integrated pest management, pp. 135139 In Hallman GJ, Denlinger DL [eds.], Temperature Sensitivity in Insects and Application in Integrated Pest Management. Westview Press, Boulder, Colorado, USA.

Kovaleski A, Ribeiro LG. 2003. Manejo de pragas na producao integrada de macas, pp. 61-76 In Protas JFS, Sanhueza RMV [eds.], Producao integrada de frutos: O caso da maca no Brasil. Embrapa Uva e Vinho, Bento Goncalves, Brazil.

Kovaleski A, Sugayama RL, Uramoto K, Malavasi A. 2000. Rio Grande do Sul, pp. 285-290 In Malavasi A, Zucchi RA [eds.], Moscas-das-frutas de importancia economica no Brasil--Conhecimento basico e aplicado. Holos Editora, Ribeirao Preto, Brazil.

Leal MR, Aguiar-Menezes EL, Filho ML, Ribeiro JCR, Menezes EB. 2008. Capacidade de sobrevivencia e dispersao de Diachasmimorpha longicaudata um parasitoide exotico de larva-de-moscas. Embrapa Agrobiologia Seropedica, Brazil.

Lira Junior JS, Bezerra JEF, Leberman IE, Silva Junior JFD. 2007. Pitangueira. Liceu, Recife, Brazil.

Lopez OP, Henaut Y, Cancino J, Lambin M, Cruz-Lopez L, Rojas JC. 2009. Is host size an indicator of quality in the mass-reared parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae)? Florida Entomologist 92: 441-449.

Manica I. 2002. Frutas nativas, silvestres e exoticas 2: tecnicas de producao e mercado: Feijoa, figo-da-India, fruta-pao, jaca, lichia e mangaba. Cinco Continentes, Porto Alegre, Brazil.

Meirelles RN, Redaelli LR, Ourique CB. 2013. Comparative biology of Diachasmimorpha longicaudata (Hymenoptera: Braconidae) reared on Anastrepha fraterculus and Ceratitis capitata (Diptera: Tephritidae). Florida Entomologist 96: 412-418.

Montoya P, Liedo P, Benrey B, Cancino J, Barrera JF, Sivinski J, Aluja M. 2000. Biological control of Anastrepha spp. (Diptera: Tephritidae) in mango orchards through augmentative releases of Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). Biological Control 18: 216-224.

Nava DE, Botton M. 2010. Biecologia e controle de Anastrepha fraterculus e Ceratitis capitata em pessegueiro. Embrapa Clima Temperado, Pelotas, Brazil.

Salles LAB. 1992. Metodologia de criacao de Anastrepha fraterculus (Wied., 1830) (Diptera: Tephritidae) em dieta artificial em laboratorio. Anais da Sociedade Entomologica do Brasil 21: 479-486.

Salles LAB. 1995. Bioecologia e controle da mosca-das-frutas sul-americana. Embrapa/CPACT, Pelotas, Brazil.

Simao S. 1971. Manual de fruticultura. Agronomia Ceres, Sao Paulo, Brazil.

Teran HR. 1977. Comportamiento alimentario y su correlacion a la reproduccion en hembras de Ceratitis capitata (Wied.) (Diptera: Tephritidae). Revista Agronomica del Noroeste Argentino 14: 17-34.

Zucchi RA. 2000. Especies de Anastrepha, sinonimias, plantas hospedeiras e parasitoides, pp. 41-48 In Malavasi A, Zucchi RA [eds.], Moscas-das-frutas de importancia economica no Brasil--Conhecimento basico e aplicado. Holos Editora, Ribeirao Preto, Brazil.

Rafael Narciso Meirelles *, Luiza Rodrigues Redaelli, and Claudia Bernardes Ourique

Programa de Pos-Graduacao em Fitotecnia, Faculdade de Agronomia, Universidade Federal do Rio Grande do Sul. Av. Bento Goncalves 7712, 91540-000. Porto Alegre, RS, Brasil

* Corresponding author; E-mail:
Table 1. Average ([+ or -] SE) development periods and their ranges
(d) of Diachasmimorpha longicaudata reared in Anastrepha fraterculus
[Dl (Af)] and in Ceratitis capitata [Dl (Cc)] larvae at 15, 21, 25,
28, and 31 [degrees]C, at 65 [+ or -] 10% RH and a photoperiod
of 14:10 h L:D.

                         Dl (Af)

([degrees]C)      Development period (d)      N      Range (d)

15                          nd               --         --
18                 39.3 [+ or -] 0.48Aa      109       33-62
21                 28.1 [+ or -] 0.31Ba      116       23-39
25                 17.9 [+ or -] 0.13Ca      114       15-23
28                 14.6 [+ or -] 0.11Da      123       13-20
31                          nd               --         --

                         Dl (Cc)

([degrees]C)      Development period (d)      N      Range (d)

15                          nd               --         --
18                 34.8 [+ or -] 0.29Ab      118       31-42
21                 25.3 [+ or -] 0.17Bb      200       22-33
25                 18.5 [+ or -] 0.15Cb      109       17-26
28                 14.8 [+ or -] 0.08Db      132       14-18
31                 15.4 [+ or -] 0.15D       109       13-19

Means followed by the same uppercase letter within a column
(Kruskal-Wallis test, P = 0.05) or lowercase letter within a
row (Mann-Whitney test, P = 0.05) are not significantly
different. N, number of observations; nd, no development.

Table 2. Lower development threshold (Dt), thermal constant (K) in
degree days (DD), linear equation of development speed (1/D), and
determination coefficient ([R.sup.2]) of the biological cycle (egg
to adult) of Diachasmimorpha longicaudata reared either in Anastrepha
fraterculus or in Ceratitis capitata at 15, 18, 21, 25, 28, and
31 [degrees]C, at 65 [+ or -] 10% RH and a photoperiod of 14:10 h L:D.

Host                  Dt         K (DD)     Regression equation
A. fraterculus       12.5        227.27    y = -0.0550 + 0.0044X
C. capitata          7.83        322.58    y = -0.0243 + 0.0031X

Host               [R.sup.2]        F         P

A. fraterculus       0.99          288     <0.0001
C. capitata          0.93         46.6      0.0064

Table 3. Average ([+ or -] SE) longevities and their ranges (d) of
Diachasmimorpha longicaudata reared either in Anastrepha fraterculus
[Dl (Af)] or in Ceratitis capitata [Dl (Cc)] at 18, 21, 25, 28, and
31 [degrees]C, at 65 [+ or -] 10% RH and a photoperiod of 14:10 h L:D.

                                      Dl (Af)

([degrees]C)           Mean (d)            N      Range (d)

18               52.7 [+ or -] 2.49Aa     109       4-112
21               29.5 [+ or -] 1.25Ba     116       6-68
25               27.0 [+ or -] 1.64Ba     108       3-76
28               9.2 [+ or -] 0.46Ca      123       1-28
31                    No adults           --         --

                                      Dl (Cc)

([degrees]C)           Mean (d)            N      Range (d)

18               53.3 [+ or -] 3.05Aa     111       2-145
21               39.4 [+ or -] 1.69Ab     120       8-107
25               29.5 [+ or -] 1.76Ba     109       1-77
28               11.3 [+ or -] 0.75Ca     111       3-34
31               11.5 [+ or -] 0.70C      109       1-30

Means followed by the same uppercase letter within a column
(Kruskal-Wallis test, P = 0.05) or lowercase letter within a row
(Mann-Whitney test, P = 0.05) are not significantly different.
N, number of observations.

Table 4. Estimated number of generations of Diachasmimorpha
longicaudata per year in 4 fruit-producing counties in Rio Grande
Do Sul, Brazil, based on annual average temperatures ([degrees]C)
and annual accumulated degree days (DD), for D. longicaudata
developing in Anastrepha fraterculus [Dl (Af)] (Dt of 12.5 [degrees]C
for the egg-to-adult period) or Ceratitis capitata [Dl (Cc)] (Dt of
7.83 [degrees]C for the egg-to-adult period).

                                               Dl (Af)

                   Annual average
                     temperature          DD             No. of
Counties            ([degrees]C)     accumulated    generations/year

Bento Goncalves         16.8           1,590.2            6.99
Porto Alegre            19.5           2,535.8            11.16
Pelotas                 17.5           1,812.2            7.97
Vacaria                 15.2           1,100.1            4.84

                              Dl (Cc)

                         DD             No. of
Counties            accumulated     generation/year

Bento Goncalves       3,241.7            10.05
Porto Alegre          4,231.8            13.12
Pelotas               3,513.3            10.89
Vacaria               2,670.2            8.28
COPYRIGHT 2015 Florida Entomological Society
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2015 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Title Annotation:Research Papers
Author:Meirelles, Rafael Narciso; Redaelli, Luiza Rodrigues; Ourique, Claudia Bernardes
Publication:Florida Entomologist
Article Type:Report
Date:Dec 1, 2015
Previous Article:Species richness of noctuid moths (Lepidoptera: Noctuidae) from the state of Guanajuato, Mexico.
Next Article:Trichoplusia ni (Lepidoptera: noctuidae) survival, immune response, and gut bacteria changes after exposure to Azadirachta indica (Sapindales:...

Terms of use | Privacy policy | Copyright © 2018 Farlex, Inc. | Feedback | For webmasters