Effect of different diets on Tyrophagus putrescentiae population and amelioration of their immunological disorder by garlic

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Nabavi, S. Arshi, A. Bahrami, Z. Aryan, M.H. Bemanian, H. Esmaeilzadeh, F. Jalali, S.B. Pousti, N. Rezaei" "autores" => array:9 [ 0 => array:2 [ "nombre" => "M." "apellidos" => "Nabavi" ] 1 => array:2 [ "nombre" => "S." "apellidos" => "Arshi" ] 2 => array:2 [ "nombre" => "A." "apellidos" => "Bahrami" ] 3 => array:2 [ "nombre" => "Z." "apellidos" => "Aryan" ] 4 => array:2 [ "nombre" => "M.H." "apellidos" => "Bemanian" ] 5 => array:2 [ "nombre" => "H." "apellidos" => "Esmaeilzadeh" ] 6 => array:2 [ "nombre" => "F." "apellidos" => "Jalali" ] 7 => array:2 [ "nombre" => "S.B." "apellidos" => "Pousti" ] 8 => array:2 [ "nombre" => "N." "apellidos" => "Rezaei" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054613001924?idApp=UINPBA00004N" "url" => "/03010546/0000004200000005/v1_201409050044/S0301054613001924/v1_201409050044/en/main.assets" ] "itemAnterior" => array:18 [ "pii" => "S0301054613001961" "issn" => "03010546" "doi" => "10.1016/j.aller.2013.05.005" "estado" => "S300" "fechaPublicacion" => "2014-09-01" "aid" => "541" "copyright" => "SEICAP" "documento" => "article" "crossmark" => 0 "subdocumento" => "fla" "cita" => "Allergol Immunopathol (Madr). 2014;42:449-58" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:2 [ "total" => 763 "formatos" => array:3 [ "EPUB" => 11 "HTML" => 486 "PDF" => 266 ] ] "en" => array:11 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Risk factors affecting asthma prevalence in adolescents living in Istanbul, Turkey" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => "en" "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "449" "paginaFinal" => "458" ] ] "contieneResumen" => array:1 [ "en" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "A. Akcay, Z. Tamay, A.B. Hocaoglu, A. Ergin, N. Guler" "autores" => array:5 [ 0 => array:2 [ "nombre" => "A." "apellidos" => "Akcay" ] 1 => array:2 [ "nombre" => "Z." "apellidos" => "Tamay" ] 2 => array:2 [ "nombre" => "A.B." "apellidos" => "Hocaoglu" ] 3 => array:2 [ "nombre" => "A." "apellidos" => "Ergin" ] 4 => array:2 [ "nombre" => "N." "apellidos" => "Guler" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054613001961?idApp=UINPBA00004N" "url" => "/03010546/0000004200000005/v1_201409050044/S0301054613001961/v1_201409050044/en/main.assets" ] "en" => array:19 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Effect of different diets on Tyrophagus putrescentiae population and amelioration of their immunological disorder by garlic" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "459" "paginaFinal" => "464" ] ] "autores" => array:1 [ 0 => array:3 [ "autoresLista" => "Bahaa K.A. Abdel-Salam" "autores" => array:1 [ 0 => array:3 [ "nombre" => "Bahaa K.A." "apellidos" => "Abdel-Salam" "email" => array:2 [ 0 => "Bahaa_2000_1999@yahoo.com" 1 => "Bahaa.abdelsalam@science.miniauniv.edu.eg" ] ] ] "afiliaciones" => array:2 [ 0 => array:2 [ "entidad" => "Zoology Department, Faculty of Science, 61519 El-Minia University, El-Minia, Egypt" "identificador" => "aff0005" ] 1 => array:2 [ "entidad" => "Biology Department, Deanship of PreparatoryYear, Shaqra University, 11961 Shaqra, Saudi Arabia" "identificador" => "aff0010" ] ] ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 2329 "Ancho" => 2333 "Tamanyo" => 1103943 ] ] "descripcion" => array:1 [ "en" => "Tyrophagus putrescentiae, (A) female whole mount; (B) male whole mount; (C) dorsal view with Sci seta longer than Sce seta; (D) ventral view of male with two anal suckers." ] ] ] "textoCompleto" => "IntroductionMedicinal plants have been used to cure human illness since ancient times. Certain types of these plants are believed to promote positive health and maintain organism resistance against infection by re-establishing body equilibrium and conditioning the body tissues. Among these plants, garlic (Allium sativum) a traditional dietary and medicinal applications as an anti-microbial agent<a class="elsevierStyleCrossRef" href="#bib0005">1</a> is a common food spice widely distributed and used in all parts of the world as a spice and herbal medicine for the prevention and treatment of a variety of diseases, ranging from infections to heart diseases.<a class="elsevierStyleCrossRef" href="#bib0010">2</a> Garlic is thought to have various pharmacological properties and medical applications. It is mainly consumed as a condiment in various prepared foods.<a class="elsevierStyleCrossRef" href="#bib0015">3</a> This prompted us to propose garlic as an asthmatic modulator medicinal plant.Understanding the nature of environmental triggers is fundamental in attempts to prevent/reduce allergic diseases. As previously noted, exposure to common aeroallergens, especially perennial inhalable allergens such as mite, is associated with a significantly increased risk for asthma.<a class="elsevierStyleCrossRef" href="#bib0020">4</a> Mite infestation is associated with negative effects on man and his resources. Storage mites consume stored grain and oil seeds, transfer toxicogenic microorganisms and produce allergens, thereby causing occupational allergies and endangering food safety.<a class="elsevierStyleCrossRef" href="#bib0025">5</a> The storage mite, Tyrophagus putrescentiae is found in stored products such as dried eggs, ham, herring meal, cheese, and different kinds of nuts.<a class="elsevierStyleCrossRef" href="#bib0030">6</a> Storage mites are the cause of occupational allergic diseases of bakers<a class="elsevierStyleCrossRef" href="#bib0035">7</a> and food industrial workers<a class="elsevierStyleCrossRefs" href="#bib0040">8,9</a> and among residents in agricultural environments.<a class="elsevierStyleCrossRef" href="#bib0050">10</a> En-Chih et al. had also mentioned that both T. putrescentiae and Dermatophagoides pteronyssinus are causative factors for the development of airway hypersensitivity.<a class="elsevierStyleCrossRef" href="#bib0055">11</a> The mass rearing processes were also reported elsewhere.<a class="elsevierStyleCrossRefs" href="#bib0060">12,13</a>The storage mite T. putrescentiae causes allergic response and IgE production in animals.<a class="elsevierStyleCrossRef" href="#bib0070">14</a> The asthma symptoms are caused by inhalation of live mites and fragments of dead mites, or their excretory pellets.<a class="elsevierStyleCrossRef" href="#bib0075">15</a>Bronchial asthma is a chronic inflammatory disease of the airway.<a class="elsevierStyleCrossRef" href="#bib0080">16</a> Chronic exposure to allergens triggers a distinct array of immunobiological and biochemical reactions that directly stimulate and induce abnormalities of air-way structure resulting in the development of clinical symptoms.<a class="elsevierStyleCrossRef" href="#bib0085">17</a> Allergic disorder symptoms are associated with high levels of serum allergen-specific IgE and eosinophilia.<a class="elsevierStyleCrossRef" href="#bib0090">18</a> In addition, both allergen exposure protocols result in immune-mediated airway inflammation defined by elevated levels of IgE, the T-helper cell 2 (TH2) cytokines IL-4 and eosinophils.<a class="elsevierStyleCrossRef" href="#bib0095">19</a> The pro-inflammatory cytokine, IFN-γ, promotes T-helper type-1 (Th1) responses, which down-regulate the Th2-like immune responses that are hallmarks of allergic diseases, including asthma.<a class="elsevierStyleCrossRef" href="#bib0100">20</a>The objective of the present investigation was to develop mass rearing techniques of this species by 13 different natural foods and to examine their immunological disorders in male albino rats. Modulation of these disorders was tested by garlic.Materials and methodsMass rearing of Tyrophagus putrescentiaeThe choice of the effective diets or feeding media is necessary for mass rearing of allergic mites, because the antigen preparation needs a very big numbers of allergic mites. Stored product samples, collected from El-Minia Governorate, were used for establishing a stock culture of T. putrescentiae. Live mites were isolated by a modified Berlese funnel with a wire screening<a class="elsevierStyleCrossRef" href="#bib0105">21</a> covered with muslin. Each sample was placed on the muslin and spread. Mites which escaped from light were received in a Petri dish. The bottom of the Petri dish is filled with a paste made of a mixture of charcoal and gypsum (2:8 parts) to construct a porous medium.<a class="elsevierStyleCrossRef" href="#bib0110">22</a> The rim of the dish is coated with a phaslene. Mites were sorted out, preserved and mounted on microscope slides for species identification.<a class="elsevierStyleCrossRefs" href="#bib0115">23,24</a>T. putrescentiae figures were captured by light microscopy at 400× and 600× magnifications.The following culture media obtained from local market were compared: (1) wheat germ, (2) roomy cheese, (3) nedo, (4) egg yolk, (5) potatoes, (6) fish meal, (7) copra, (8) egg albumin, (9) chicken bouillon, (10) pollens, (11) powdered bean, (12) human hair and scales, and (13) yeast extract.Five males and five females in copulation state were caged with the tested diet in certain plastic vials (5mm diam.×20mm length), cigarette paper held in place by a snap-on plastic ring confined the mites and allowed gas exchange between the culture environment and a humidity chamber environment. The bottom of each vial was filled with the last described paste. Each combination of food and species of mites was replicated five times within a test. The Petri dishes of the stock culture and the plastic vials of the tested diets were carried out at 25±1°C and 75% relative humidity (RH). The temperature was adjusted at 25°C by placing the entire system inside an incubator. The relative humidity inside the desiccators was maintained at 75% by the concentration of the KOH (22.25g of KOH per 100ml dis. water). The concentration is also sufficient to absorb the CO2 as soon as it is released.<a class="elsevierStyleCrossRef" href="#bib0125">25</a> Every two months the stock cultures were transplanted into fresh media to prevent outgrowth and a consequent breakdown of the population. Contamination with fungi was controlled by: (1) sterilising the tested diets and containers using dry heat (roughly 90°C for 30min). (2) Adding diets in small crumbled amounts. (3) Using of 75% RH instead of 80%, the deal RH needed for T. putrescentiae culture. The procedure of mite counts was as mentioned by Ree and Lee.<a class="elsevierStyleCrossRef" href="#bib0130">26</a>Crude extractsWhen the population density was sufficiently high, T. putrescentiae crude faeces were isolated by the Berlese–Tullgren method by photonegative live mites escaping. Five grams from these faecal pellets were used for each inhalation treatment. Garlic cloves were purchased from the local market in El-Minia Governorate, Egypt. They were then cleaned of any adhered dried material. Each animal was allowed free access to food 50mg of seeds and cloves daily during the course of the treatment, after fasting for about 12h.Experimental animalsMale albino rats, Rattus norvegicus (6–8 weeks old, weight 100–120g) were purchased from the Biological Supply Center, Theodore Bilharz Research Institute, TBRI, Cairo, Egypt and housed under specific pathogen-free conditions and maintained on a 12-h light-dark cycle, with food and water ad libitum. The experiments were conducted according to the ethical norm approved by the Institutional Animal Ethics Committee (IAEC). Animals were classified into four groups (10 animals each). The first group (control) was untreated. The second group was allowed free access to food 50mg of garlic daily during the course of the treatment, after fasting for about 12h. The third group was intranasal inhaled with T. putrescentiae faeces extract (5g) daily for 10 consecutive days. The fourth group inhaled as in the third group and allowed free access to food as in the second group.Cytokine and immunoglobulin in serum, 72h after the last treatment, ten animals from each group were sacrificed under chloroform anaesthesia. Blood samples were taken and centrifuged at 3000rpm for 30min. Sera were removed and kept at −20°C for the estimation of the cytokines (IFN-γ and IL-4) and IgE antibody levels by ELISA using kits purchased from R&D Systems (Minneapolis, MN, USA).Statistical analysisData were analysed using SPSS program version 13.0. Statistical analysis of the obtained data was performed using one way analysis of variance (ANOVA) test followed by least square differences (LSD) analysis for comparison between means. Results were expressed as mean±standard error (SE). Values of P<0.05 were considered statistically significant, while values of P>0.05 were considered statistically non-significant.ResultsMass rearing of Tyrophagus putrescentiaeThis species belongs to the family Acaridae. Its dorsal Setae Sci is longer than Sce. In addition, the femur of male forelegs is not enlarged and without a ventral conical process. Females do not have anal suckers that are present in males (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>).<elsevierMultimedia ident="fig0005"></elsevierMultimedia>Among 13 different media tested for mass rearing of T. putrescentiae mites, statistical analysis of the obtained data showed a significant difference (P<0.05) in mite population recorded by wheat germ, roomy cheese, nido, egg yolk and potatoes media compared to those recorded by fish meal, copra, egg albumin, chicken stocks, pollens, bean, human hair and scales and yeast extract (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>).<elsevierMultimedia ident="fig0010"></elsevierMultimedia>Quantification of INF-γThe INF-γ ELISA value demonstrated that the garlic-treated group (1556.47±179.452pg/ml) was lower than the control group (1885.88±003.867pg/ml). The groups treated with T. putrescentiae faeces produced by wheat germ (8030.29±422.930pg/ml), roomy cheese (8113.06±036.869pg/ml), nido (7554.91±186.849pg/ml), egg yolk (7430.97±214.261pg/ml) and potatoes (6748.63±102.913pg/ml) were higher than the groups treated with garlic in addition to the above five diets (4201.97±141.100pg/ml, 4101.43±110.254pg/ml, 4129.77±147.799pg/ml, 3275.37±38.531pg/ml, 3022.40±105.782pg/ml, respectively) (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>). This study also showed that there is a significant difference in INF-γ level between garlic-treated group and either the native or faeces-treated group (P<0.05).<elsevierMultimedia ident="fig0015"></elsevierMultimedia>Quantification of IL-4Results of ELISA in <a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a> showed that IL-4 recorded 567.07±07.929pg/ml in control group. The inhaled group with T. putrescentiae faeces counted high level of IL-4 (812.51±22.897pg/ml). In contrast, the groups treated with T. putrescentiae faeces produced by wheat germ (231.12±03.448pg/ml), roomy cheese (196.90±05.448pg/ml), nido (187.43±05.339pg/ml), egg yolk (154.51±10.097pg/ml) and potatoes (150.80±08.091pg/ml) are lower than the groups treated with garlic in addition to the above five diets (674.22±13.834pg/ml, 645.90±15.277pg/ml, 640.54±13.643pg/ml, 532.48±15.371pg/ml, 489.12±17.354pg/ml, respectively) (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>). Statistical analysis showed a significant difference in IL-4 between garlic-treated group and either control or faeces-treated group, P<0.05.<elsevierMultimedia ident="fig0020"></elsevierMultimedia>Quantification of IgEIn contrast to IL-4, ELISA tests of IgE showed that the garlic-treated group (36.30±0.427ng/ml) is lower than the control group (44.28±0.400) and the groups treated with T. putrescentiae faeces produced by wheat germ (112.87±2.798pg/ml), roomy cheese (77.71±7.646pg/ml), nido (78.46±5.449pg/ml), egg yolk (61.90±4.374pg/ml) and potatoes (58.84±3.213pg/ml) are higher than the groups treated with garlic in addition to the above five diets (49.51±3.642pg/ml, 51.91±1.954pg/ml, 49.87±1.691pg/ml, 47.65±1.241pg/ml, 44.19±1.525pg/ml, respectively) (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>). Statistical analysis showed that there is a significant difference between the garlic-treated group and either the control or faeces-treated group (P<0.05).<elsevierMultimedia ident="fig0025"></elsevierMultimedia>DiscussionT. putrescentae mites are known to cause hypersensitivity reactions and asthma.<a class="elsevierStyleCrossRef" href="#bib0050">10</a> Selection of the culture media is one of the very important factors for successful mass rearing of house dust/storage mites.<a class="elsevierStyleCrossRef" href="#bib0130">26</a> The mite problems described above can have severe effects on the health.<a class="elsevierStyleCrossRef" href="#bib0075">15</a> The following points must be taken carefully to prepare a specific vaccine against house dust mite bronchial asthma, perennial rhinitis or allergic dermatitis. These are: (i) identification of the local house dust mites, (ii) determination of the sensitising mite allergen and this necessitates isolation of the identified house dust mites extracts, and (iii) identification of the specific house dust mite extract to which patients give positive reaction.Mass rearing of T. putrescentiae was carried out, since this species has proven to be the second most important allergic mite in Egypt<a class="elsevierStyleCrossRef" href="#bib0110">22</a> as well as in other places such as Sydney and New Guinea.<a class="elsevierStyleCrossRef" href="#bib0135">27</a> It was also recorded in the three governorates with high numbers. In this study the best diet was the wheat germ by which 1881 mites were gained. The second effective diet (cake) gave 1575.9 mites. Roomy cheese gave 1539.7 mites. Also nido, egg yolk and egg albumin gave a number of mites greater than one thousand. On the other hand, potatoes, fish meal, copra, chicken bouillon, pollens, powdered bean, human hair and scales and yeast extract gave a number of mites lower than 500 mites.Similar and other diets such as powdered animal food;<a class="elsevierStyleCrossRef" href="#bib0140">28</a> (yeast, clover seed, ground nuts, linseed, maize germ, peas and rice);<a class="elsevierStyleCrossRef" href="#bib0145">29</a> (mixture of ground and dried oil liver);<a class="elsevierStyleCrossRef" href="#bib0150">30</a> a mixture of human skin scales and yeast, monosaccharides, disaccharides and polysaccharides;<a class="elsevierStyleCrossRef" href="#bib0155">31</a> wheat germ;<a class="elsevierStyleCrossRef" href="#bib0135">27</a> and a mixture of roomy cheese, dried egg yolk and yeast.<a class="elsevierStyleCrossRef" href="#bib0110">22</a> Green and Woodcock mentioned that the type of food ingested can markedly affect the fecundity, duration of development, maturity, longevity and age structure of a T. putrescentiae population.<a class="elsevierStyleCrossRef" href="#bib0135">27</a>In addition to the mite mass rearing, we have investigated whether the faecal antigen is important in the development of asthma or not, where mite faecal pellets break up into many small pieces and combine with dust particles to form an allergic dust.<a class="elsevierStyleCrossRef" href="#bib0160">32</a>The potential of certain pollutants to elicit allergic airway disease has been established in both experimental models and in humans.<a class="elsevierStyleCrossRef" href="#bib0165">33</a> Short exposure (10 days) to HDM alone or to HDM and OVA resulted in equivalent levels of airway inflammation due to a robust inflammatory response in the lung. Moreover, it has been recently shown that intranasal administration of HDM daily for 10 days induces allergic sensitisation and Th2 airway inflammation, both of which are substantially reduced by concurrent treatment with anti-M-CSF antibodies.<a class="elsevierStyleCrossRef" href="#bib0170">34</a> This prompted us to expose the tested animals for about 10 days.Allergic disorders have been rising in recent years. It has been demonstrated that IgE antibodies play an important role in mediating type I hypersensitivity in humans.<a class="elsevierStyleCrossRef" href="#bib0175">35</a> In both food and inhalant allergy it is accepted that food- or HDM-specific IgE binds to high-affinity Fc¿RI on mast cells, basophiles, macrophages, and dendritic cells, as well as to low-affinity Fc¿RII on macrophages, monocytes, lymphocytes, eosinophils, and platelets.<a class="elsevierStyleCrossRef" href="#bib0180">36</a> When mite allergens penetrate mucosal barriers of the gastrointestinal or respiratory tract and contact IgE antibodies bound to mast cells or basophiles, histamine and other mediators that induce symptoms of immediate hypersensitivity are released.<a class="elsevierStyleCrossRef" href="#bib0185">37</a> Other studies have also suggested that cleavage of CD23 (low-affinity IgE receptor) may promote and enhance an IgE immune response.<a class="elsevierStyleCrossRef" href="#bib0190">38</a>Another factor to consider is cytokine production. Indeed, it has been suggested that HDM may privilege the generation of a Th2-polarised response by modulating the balance between IL-4 and IFN-γ.<a class="elsevierStyleCrossRef" href="#bib0195">39</a> In addition, various dust mite allergens act on bronchial epithelial cells in vitro to elicit the production of a number of cytokines.<a class="elsevierStyleCrossRef" href="#bib0200">40</a>IL-4 is a key anti-inflammatory cytokine and plays an important role in the cytokine network and in the Th1/Th2 balance. It is generated by helper T cells, and possibly in the early adaptive immune response by mast cells and basophils.<a class="elsevierStyleCrossRef" href="#bib0175">35</a> In this study, the low level of IL-4 in the faeces-treated group may be due to the negative feedback that is exerted by histamine itself to inhibit IL-4.<a class="elsevierStyleCrossRef" href="#bib0185">37</a>Thus, this study tended to measure the levels of INF-γ, L-4 and IgE that are involved in allergic disease. Repeated exposure to T. putrescentiae allergic faeces caused an increase in both INF-γ and IgE, and a decrease in IL-4. The high level of both INF-γ and IgE, and the low level of IL-4 may participate in airway disorder. After garlic feeding, the increase of IL-4 and the decrease of both INF-γ and IgE were maintained, and this may play an important role in the airway remodelling processes. The immunomodulatory effects on INF-γ, IL-4 and IgE balance were tested by others.<a class="elsevierStyleCrossRefs" href="#bib0070">14,41</a>Garlic in small amounts applied as a food condiment does not usually pose an important risk, except for rare specific allergic reactions in hypersensitive people.<a class="elsevierStyleCrossRef" href="#bib0210">42</a> In contrast, this study showed that some allergic disorder could be modulated by garlic.In conclusion, these data demonstrated that wheat germ, roomy cheese, nido, egg yolk and potatoes media increase the mass rearing of T. putrescentiae compared to those recorded by fish meal, copra, egg albumin, chicken stocks, pollens, bean, human hair & scales and yeast extract. Repeated exposure to T. putrescentiae allergic faeces caused an increase in both the pro-inflammatory cytokine INF-γ and the main allergic immunoglobulin IgE, and a decrease in the anti-inflammatory cytokine IL-4. The high level of both INF-γ and IgE, and the low level of IL-4 may participate in airway disorder. After garlic treating, the increase of IL-4 and the decrease of both INF-γ and IgE were maintained, and this means that garlic may play an important role in the amelioration of the airway immune response.Ethical disclosuresProtection of human and animal subjectsThe procedures followed were in accordance with the regulations of the responsible Clinical Research Ethics Committee and in accordance with those of the World Medical Association and the Helsinki Declaration.Confidentiality of dataNo patient data appears in this article.Right to privacy and informed consentNo patient data appears in this article.Conflict of interestThe author has no conflict of interest to declare." "textoCompletoSecciones" => array:1 [ "secciones" => array:10 [ 0 => array:2 [ "identificador" => "xres368090" "titulo" => array:6 [ 0 => "Abstract" 1 => "Background" 2 => "Objective" 3 => "Methods" 4 => "Results" 5 => "Conclusions" ] ] 1 => array:2 [ "identificador" => "xpalclavsec347265" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 3 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Mass rearing of Tyrophagus putrescentiae" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Crude extracts" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Experimental animals" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "Statistical analysis" ] ] ] 4 => array:3 [ "identificador" => "sec0035" "titulo" => "Results" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "sec0040" "titulo" => "Mass rearing of Tyrophagus putrescentiae" ] 1 => array:2 [ "identificador" => "sec0045" "titulo" => "Quantification of INF-γ" ] 2 => array:2 [ "identificador" => "sec0050" "titulo" => "Quantification of IL-4" ] 3 => array:2 [ "identificador" => "sec0055" "titulo" => "Quantification of IgE" ] ] ] 5 => array:2 [ "identificador" => "sec0060" "titulo" => "Discussion" ] 6 => array:3 [ "identificador" => "sec0065" "titulo" => "Ethical disclosures" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0070" "titulo" => "Protection of human and animal subjects" ] 1 => array:2 [ "identificador" => "sec0075" "titulo" => "Confidentiality of data" ] 2 => array:2 [ "identificador" => "sec0080" "titulo" => "Right to privacy and informed consent" ] ] ] 7 => array:2 [ "identificador" => "sec0085" "titulo" => "Conflict of interest" ] 8 => array:2 [ "identificador" => "xack92167" "titulo" => "Acknowledgements" ] 9 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2013-03-09" "fechaAceptado" => "2013-05-10" "PalabrasClave" => array:1 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec347265" "palabras" => array:7 [ 0 => "Mass rearing" 1 => "Natural diet" 2 => "T. putrescentiae faeces" 3 => "Allergic immune response" 4 => "IgE" 5 => "IL-4" 6 => "INF-γ" ] ] ] ] "tieneResumen" => true "resumen" => array:1 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "BackgroundThe storage mite, Tyrophagus putrescentiae, detected in the samples collected from stored products and house dust, is one of the major causes of allergic disorders. ObjectiveThe purpose of this study was to ameliorate the T. putrescentiae faeces allergic immunological disorder by garlic. MethodsAlbino experimental rats were classified into control, inhaled and treated groups. Mass rearing of T. putrescentiae on different diets, and ELISA of some cytokines and IgE techniques were used. ResultsThe results obtained showed the highest population of T. putrescentiae reared in four from thirteen tested diets. In addition, significantly higher serum levels of INF-γ and IgE were found in rats treated with faeces than the other groups; especially the garlic-treated group. In contrast, IL-4 was lower in faeces-treated rats than the others; however, the control group had the highest level of IL-4. Statistical analysis of data showed a significant difference between the garlic-treated group and either control or faeces-treated groups (P<0.05). ConclusionsThe population of T. putrescentiae mites peaked in four from thirteen tested diets. The immunological disorder caused by repeated exposure to T. putrescentiae faeces might be modulated by garlic." ] ] "multimedia" => array:5 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 2329 "Ancho" => 2333 "Tamanyo" => 1103943 ] ] "descripcion" => array:1 [ "en" => "Tyrophagus putrescentiae, (A) female whole mount; (B) male whole mount; (C) dorsal view with Sci seta longer than Sce seta; (D) ventral view of male with two anal suckers." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1000 "Ancho" => 1594 "Tamanyo" => 109110 ] ] "descripcion" => array:1 [ "en" => "Comparative mass rearing of T. putrescentiae under different culture media for two months." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 1045 "Ancho" => 1609 "Tamanyo" => 124959 ] ] "descripcion" => array:1 [ "en" => "Changes in INF-γ levels in male albino rats inhaled with T. putrescentiae mite faeces and treated with garlic." ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 1042 "Ancho" => 1578 "Tamanyo" => 120644 ] ] "descripcion" => array:1 [ "en" => "Changes in IL-4 levels in male albino rats inhaled with T. putrescentiae mite faeces and treated with garlic." ] ] 4 => array:7 [ "identificador" => "fig0025" "etiqueta" => "Figure 5" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr5.jpeg" "Alto" => 1005 "Ancho" => 1587 "Tamanyo" => 109495 ] ] "descripcion" => array:1 [ "en" => "Changes in IgE levels in male albino rats inhaled with T. putrescentiae mite faeces and treated with garlic." ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:42 [ 0 => array:3 [ "identificador" => "bib0005" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Antimicrobial properties of garlic oil against human enteric bacteria: evaluation of methodologies and comparisons with garlic oil sulfides and garlic powder" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ 0 => "Z.M. 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"vista" => "all" ] ] ] "idiomaDefecto" => "en" "url" => "/03010546/0000004200000005/v1_201409050044/S0301054613001456/v1_201409050044/en/main.assets" "Apartado" => array:4 [ "identificador" => "5554" "tipo" => "SECCION" "en" => array:2 [ "titulo" => "Original articles" "idiomaDefecto" => true ] "idiomaDefecto" => "en" ] "PDF" => "https://static.elsevier.es/multimedia/03010546/0000004200000005/v1_201409050044/S0301054613001456/v1_201409050044/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054613001456?idApp=UINPBA00004N"]

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Bahaa K.A. Abdel-Salam

Zoology Department, Faculty of Science, 61519 El-Minia University, El-Minia, Egypt

Biology Department, Deanship of PreparatoryYear, Shaqra University, 11961 Shaqra, Saudi Arabia

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          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">Tyrophagus putrescentiae</span>&#44; &#40;A&#41; female whole mount&#59; &#40;B&#41; male whole mount&#59; &#40;C&#41; dorsal view with Sci seta longer than Sce seta&#59; &#40;D&#41; ventral view of male with two anal suckers&#46;</p>"
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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Medicinal plants have been used to cure human illness since ancient times&#46; Certain types of these plants are believed to promote positive health and maintain organism resistance against infection by re-establishing body equilibrium and conditioning the body tissues&#46; Among these plants&#44; garlic &#40;<span class="elsevierStyleItalic">Allium sativum</span>&#41; a traditional dietary and medicinal applications as an anti-microbial agent<a class="elsevierStyleCrossRef" href="#bib0005"><span class="elsevierStyleSup">1</span></a> is a common food spice widely distributed and used in all parts of the world as a spice and herbal medicine for the prevention and treatment of a variety of diseases&#44; ranging from infections to heart diseases&#46;<a class="elsevierStyleCrossRef" href="#bib0010"><span class="elsevierStyleSup">2</span></a> Garlic is thought to have various pharmacological properties and medical applications&#46; It is mainly consumed as a condiment in various prepared foods&#46;<a class="elsevierStyleCrossRef" href="#bib0015"><span class="elsevierStyleSup">3</span></a> This prompted us to propose garlic as an asthmatic modulator medicinal plant&#46;</p><p id="par0010" class="elsevierStylePara elsevierViewall">Understanding the nature of environmental triggers is fundamental in attempts to prevent&#47;reduce allergic diseases&#46; As previously noted&#44; exposure to common aeroallergens&#44; especially perennial inhalable allergens such as mite&#44; is associated with a significantly increased risk for asthma&#46;<a class="elsevierStyleCrossRef" href="#bib0020"><span class="elsevierStyleSup">4</span></a> Mite infestation is associated with negative effects on man and his resources&#46; Storage mites consume stored grain and oil seeds&#44; transfer toxicogenic microorganisms and produce allergens&#44; thereby causing occupational allergies and endangering food safety&#46;<a class="elsevierStyleCrossRef" href="#bib0025"><span class="elsevierStyleSup">5</span></a> The storage mite&#44; <span class="elsevierStyleItalic">Tyrophagus putrescentiae</span> is found in stored products such as dried eggs&#44; ham&#44; herring meal&#44; cheese&#44; and different kinds of nuts&#46;<a class="elsevierStyleCrossRef" href="#bib0030"><span class="elsevierStyleSup">6</span></a> Storage mites are the cause of occupational allergic diseases of bakers<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> and food industrial workers<a class="elsevierStyleCrossRefs" href="#bib0040"><span class="elsevierStyleSup">8&#44;9</span></a> and among residents in agricultural environments&#46;<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">10</span></a> En-Chih et al&#46; had also mentioned that both <span class="elsevierStyleItalic">T&#46; putrescentiae</span> and <span class="elsevierStyleItalic">Dermatophagoides pteronyssinus</span> are causative factors for the development of airway hypersensitivity&#46;<a class="elsevierStyleCrossRef" href="#bib0055"><span class="elsevierStyleSup">11</span></a> The mass rearing processes were also reported elsewhere&#46;<a class="elsevierStyleCrossRefs" href="#bib0060"><span class="elsevierStyleSup">12&#44;13</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">The storage mite <span class="elsevierStyleItalic">T&#46; putrescentiae</span> causes allergic response and IgE production in animals&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a> The asthma symptoms are caused by inhalation of live mites and fragments of dead mites&#44; or their excretory pellets&#46;<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">15</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">Bronchial asthma is a chronic inflammatory disease of the airway&#46;<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">16</span></a> Chronic exposure to allergens triggers a distinct array of immunobiological and biochemical reactions that directly stimulate and induce abnormalities of air-way structure resulting in the development of clinical symptoms&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a> Allergic disorder symptoms are associated with high levels of serum allergen-specific IgE and eosinophilia&#46;<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">18</span></a> In addition&#44; both allergen exposure protocols result in immune-mediated airway inflammation defined by elevated levels of IgE&#44; the T-helper cell 2 &#40;TH2&#41; cytokines IL-4 and eosinophils&#46;<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">19</span></a> The pro-inflammatory cytokine&#44; IFN-&#947;&#44; promotes T-helper type-1 &#40;Th1&#41; responses&#44; which down-regulate the Th2-like immune responses that are hallmarks of allergic diseases&#44; including asthma&#46;<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">20</span></a></p><p id="par0025" class="elsevierStylePara elsevierViewall">The objective of the present investigation was to develop mass rearing techniques of this species by 13 different natural foods and to examine their immunological disorders in male albino rats&#46; Modulation of these disorders was tested by garlic&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Mass rearing of <span class="elsevierStyleItalic">Tyrophagus putrescentiae</span></span><p id="par0030" class="elsevierStylePara elsevierViewall">The choice of the effective diets or feeding media is necessary for mass rearing of allergic mites&#44; because the antigen preparation needs a very big numbers of allergic mites&#46; Stored product samples&#44; collected from El-Minia Governorate&#44; were used for establishing a stock culture of <span class="elsevierStyleItalic">T&#46; putrescentiae</span>&#46; Live mites were isolated by a modified Berlese funnel with a wire screening<a class="elsevierStyleCrossRef" href="#bib0105"><span class="elsevierStyleSup">21</span></a> covered with muslin&#46; Each sample was placed on the muslin and spread&#46; Mites which escaped from light were received in a Petri dish&#46; The bottom of the Petri dish is filled with a paste made of a mixture of charcoal and gypsum &#40;2&#58;8 parts&#41; to construct a porous medium&#46;<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">22</span></a> The rim of the dish is coated with a phaslene&#46; Mites were sorted out&#44; preserved and mounted on microscope slides for species identification&#46;<a class="elsevierStyleCrossRefs" href="#bib0115"><span class="elsevierStyleSup">23&#44;24</span></a><span class="elsevierStyleItalic">T&#46; putrescentiae</span> figures were captured by light microscopy at 400&#215; and 600&#215; magnifications&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">The following culture media obtained from local market were compared&#58; &#40;1&#41; wheat germ&#44; &#40;2&#41; roomy cheese&#44; &#40;3&#41; nedo&#44; &#40;4&#41; egg yolk&#44; &#40;5&#41; potatoes&#44; &#40;6&#41; fish meal&#44; &#40;7&#41; copra&#44; &#40;8&#41; egg albumin&#44; &#40;9&#41; chicken bouillon&#44; &#40;10&#41; pollens&#44; &#40;11&#41; powdered bean&#44; &#40;12&#41; human hair and scales&#44; and &#40;13&#41; yeast extract&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">Five males and five females in copulation state were caged with the tested diet in certain plastic vials &#40;5<span class="elsevierStyleHsp" style=""></span>mm diam&#46;<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>20<span class="elsevierStyleHsp" style=""></span>mm length&#41;&#44; cigarette paper held in place by a snap-on plastic ring confined the mites and allowed gas exchange between the culture environment and a humidity chamber environment&#46; The bottom of each vial was filled with the last described paste&#46; Each combination of food and species of mites was replicated five times within a test&#46; The Petri dishes of the stock culture and the plastic vials of the tested diets were carried out at 25<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1<span class="elsevierStyleHsp" style=""></span>&#176;C and 75&#37; relative humidity &#40;RH&#41;&#46; The temperature was adjusted at 25<span class="elsevierStyleHsp" style=""></span>&#176;C by placing the entire system inside an incubator&#46; The relative humidity inside the desiccators was maintained at 75&#37; by the concentration of the KOH &#40;22&#46;25<span class="elsevierStyleHsp" style=""></span>g of KOH per 100<span class="elsevierStyleHsp" style=""></span>ml dis&#46; water&#41;&#46; The concentration is also sufficient to absorb the CO<span class="elsevierStyleInf">2</span> as soon as it is released&#46;<a class="elsevierStyleCrossRef" href="#bib0125"><span class="elsevierStyleSup">25</span></a> Every two months the stock cultures were transplanted into fresh media to prevent outgrowth and a consequent breakdown of the population&#46; Contamination with fungi was controlled by&#58; &#40;1&#41; sterilising the tested diets and containers using dry heat &#40;roughly 90<span class="elsevierStyleHsp" style=""></span>&#176;C for 30<span class="elsevierStyleHsp" style=""></span>min&#41;&#46; &#40;2&#41; Adding diets in small crumbled amounts&#46; &#40;3&#41; Using of 75&#37; RH instead of 80&#37;&#44; the deal RH needed for <span class="elsevierStyleItalic">T&#46; putrescentiae</span> culture&#46; The procedure of mite counts was as mentioned by Ree and Lee&#46;<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">26</span></a></p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Crude extracts</span><p id="par0045" class="elsevierStylePara elsevierViewall">When the population density was sufficiently high&#44; <span class="elsevierStyleItalic">T&#46; putrescentiae</span> crude faeces were isolated by the Berlese&#8211;Tullgren method by photonegative live mites escaping&#46; Five grams from these faecal pellets were used for each inhalation treatment&#46; Garlic cloves were purchased from the local market in El-Minia Governorate&#44; Egypt&#46; They were then cleaned of any adhered dried material&#46; Each animal was allowed free access to food 50<span class="elsevierStyleHsp" style=""></span>mg of seeds and cloves daily during the course of the treatment&#44; after fasting for about 12<span class="elsevierStyleHsp" style=""></span>h&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Experimental animals</span><p id="par0050" class="elsevierStylePara elsevierViewall">Male albino rats&#44; <span class="elsevierStyleItalic">Rattus norvegicus</span> &#40;6&#8211;8 weeks old&#44; weight 100&#8211;120<span class="elsevierStyleHsp" style=""></span>g&#41; were purchased from the Biological Supply Center&#44; Theodore Bilharz Research Institute&#44; TBRI&#44; Cairo&#44; Egypt and housed under specific pathogen-free conditions and maintained on a 12-h light-dark cycle&#44; with food and water ad libitum&#46; The experiments were conducted according to the ethical norm approved by the Institutional Animal Ethics Committee &#40;IAEC&#41;&#46; Animals were classified into four groups &#40;10 animals each&#41;&#46; The first group &#40;control&#41; was untreated&#46; The second group was allowed free access to food 50<span class="elsevierStyleHsp" style=""></span>mg of garlic daily during the course of the treatment&#44; after fasting for about 12<span class="elsevierStyleHsp" style=""></span>h&#46; The third group was intranasal inhaled with <span class="elsevierStyleItalic">T&#46; putrescentiae</span> faeces extract &#40;5<span class="elsevierStyleHsp" style=""></span>g&#41; daily for 10 consecutive days&#46; The fourth group inhaled as in the third group and allowed free access to food as in the second group&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">Cytokine and immunoglobulin in serum&#44; 72<span class="elsevierStyleHsp" style=""></span>h after the last treatment&#44; ten animals from each group were sacrificed under chloroform anaesthesia&#46; Blood samples were taken and centrifuged at 3000<span class="elsevierStyleHsp" style=""></span>rpm for 30<span class="elsevierStyleHsp" style=""></span>min&#46; Sera were removed and kept at &#8722;20<span class="elsevierStyleHsp" style=""></span>&#176;C for the estimation of the cytokines &#40;IFN-&#947; and IL-4&#41; and IgE antibody levels by ELISA using kits purchased from R&#38;D Systems &#40;Minneapolis&#44; MN&#44; USA&#41;&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Statistical analysis</span><p id="par0060" class="elsevierStylePara elsevierViewall">Data were analysed using SPSS program version 13&#46;0&#46; Statistical analysis of the obtained data was performed using one way analysis of variance &#40;ANOVA&#41; test followed by least square differences &#40;LSD&#41; analysis for comparison between means&#46; Results were expressed as mean<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard error &#40;SE&#41;&#46; Values of <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 were considered statistically significant&#44; while values of <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>0&#46;05 were considered statistically non-significant&#46;</p></span></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Results</span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Mass rearing of <span class="elsevierStyleItalic">Tyrophagus putrescentiae</span></span><p id="par0065" class="elsevierStylePara elsevierViewall">This species belongs to the family Acaridae&#46; 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roomy cheese &#40;8113&#46;06<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>036&#46;869<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; nido &#40;7554&#46;91<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>186&#46;849<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; egg yolk &#40;7430&#46;97<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>214&#46;261<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; and potatoes &#40;6748&#46;63<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>102&#46;913<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; were higher than the groups treated with garlic in addition to the above five diets &#40;4201&#46;97<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>141&#46;100<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 4101&#46;43<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>110&#46;254<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 4129&#46;77<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>147&#46;799<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 3275&#46;37<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>38&#46;531<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 3022&#46;40<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>105&#46;782<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; respectively&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>&#41;&#46; This study also showed that there is a significant difference in INF-&#947; level between garlic-treated group and either the native or faeces-treated group &#40;<span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Quantification of IL-4</span><p id="par0080" class="elsevierStylePara elsevierViewall">Results of ELISA in <a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a> showed that IL-4 recorded 567&#46;07<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>07&#46;929<span class="elsevierStyleHsp" style=""></span>pg&#47;ml in control group&#46; The inhaled group with <span class="elsevierStyleItalic">T&#46; putrescentiae</span> faeces counted high level of IL-4 &#40;812&#46;51<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>22&#46;897<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#46; In contrast&#44; the groups treated with <span class="elsevierStyleItalic">T&#46; putrescentiae</span> faeces produced by wheat germ &#40;231&#46;12<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>03&#46;448<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; roomy cheese &#40;196&#46;90<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>05&#46;448<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; nido &#40;187&#46;43<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>05&#46;339<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; egg yolk &#40;154&#46;51<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>10&#46;097<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; and potatoes &#40;150&#46;80<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>08&#46;091<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; are lower than the groups treated with garlic in addition to the above five diets &#40;674&#46;22<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>13&#46;834<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 645&#46;90<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>15&#46;277<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 640&#46;54<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>13&#46;643<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 532&#46;48<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>15&#46;371<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 489&#46;12<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>17&#46;354<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; respectively&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>&#41;&#46; Statistical analysis showed a significant difference in IL-4 between garlic-treated group and either control or faeces-treated group&#44; <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#46;</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Quantification of IgE</span><p id="par0085" class="elsevierStylePara elsevierViewall">In contrast to IL-4&#44; ELISA tests of IgE showed that the garlic-treated group &#40;36&#46;30<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;427<span class="elsevierStyleHsp" style=""></span>ng&#47;ml&#41; is lower than the control group &#40;44&#46;28<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;400&#41; and the groups treated with <span class="elsevierStyleItalic">T&#46; putrescentiae</span> faeces produced by wheat germ &#40;112&#46;87<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>2&#46;798<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; roomy cheese &#40;77&#46;71<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>7&#46;646<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; nido &#40;78&#46;46<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>5&#46;449<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; egg yolk &#40;61&#46;90<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>4&#46;374<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; and potatoes &#40;58&#46;84<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;213<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; are higher than the groups treated with garlic in addition to the above five diets &#40;49&#46;51<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;642<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 51&#46;91<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;954<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 49&#46;87<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;691<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 47&#46;65<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;241<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 44&#46;19<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;525<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; respectively&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>&#41;&#46; Statistical analysis showed that there is a significant difference between the garlic-treated group and either the control or faeces-treated group &#40;<span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46;</p><elsevierMultimedia ident="fig0025"></elsevierMultimedia></span></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Discussion</span><p id="par0090" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">T&#46; putrescentae</span> mites are known to cause hypersensitivity reactions and asthma&#46;<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">10</span></a> Selection of the culture media is one of the very important factors for successful mass rearing of house dust&#47;storage mites&#46;<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">26</span></a> The mite problems described above can have severe effects on the health&#46;<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">15</span></a> The following points must be taken carefully to prepare a specific vaccine against house dust mite bronchial asthma&#44; perennial rhinitis or allergic dermatitis&#46; These are&#58; &#40;i&#41; identification of the local house dust mites&#44; &#40;ii&#41; determination of the sensitising mite allergen and this necessitates isolation of the identified house dust mites extracts&#44; and &#40;iii&#41; identification of the specific house dust mite extract to which patients give positive reaction&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">Mass rearing of <span class="elsevierStyleItalic">T&#46; putrescentiae</span> was carried out&#44; since this species has proven to be the second most important allergic mite in Egypt<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">22</span></a> as well as in other places such as Sydney and New Guinea&#46;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">27</span></a> It was also recorded in the three governorates with high numbers&#46; In this study the best diet was the wheat germ by which 1881 mites were gained&#46; The second effective diet &#40;cake&#41; gave 1575&#46;9 mites&#46; Roomy cheese gave 1539&#46;7 mites&#46; Also nido&#44; egg yolk and egg albumin gave a number of mites greater than one thousand&#46; On the other hand&#44; potatoes&#44; fish meal&#44; copra&#44; chicken bouillon&#44; pollens&#44; powdered bean&#44; human hair and scales and yeast extract gave a number of mites lower than 500 mites&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">Similar and other diets such as powdered animal food&#59;<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">28</span></a> &#40;yeast&#44; clover seed&#44; ground nuts&#44; linseed&#44; maize germ&#44; peas and rice&#41;&#59;<a class="elsevierStyleCrossRef" href="#bib0145"><span class="elsevierStyleSup">29</span></a> &#40;mixture of ground and dried oil liver&#41;&#59;<a class="elsevierStyleCrossRef" href="#bib0150"><span class="elsevierStyleSup">30</span></a> a mixture of human skin scales and yeast&#44; monosaccharides&#44; disaccharides and polysaccharides&#59;<a class="elsevierStyleCrossRef" href="#bib0155"><span class="elsevierStyleSup">31</span></a> wheat germ&#59;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">27</span></a> and a mixture of roomy cheese&#44; dried egg yolk and yeast&#46;<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">22</span></a> Green and Woodcock mentioned that the type of food ingested can markedly affect the fecundity&#44; duration of development&#44; maturity&#44; longevity and age structure of a <span class="elsevierStyleItalic">T&#46; putrescentiae</span> population&#46;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">27</span></a></p><p id="par0105" class="elsevierStylePara elsevierViewall">In addition to the mite mass rearing&#44; we have investigated whether the faecal antigen is important in the development of asthma or not&#44; where mite faecal pellets break up into many small pieces and combine with dust particles to form an allergic dust&#46;<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">32</span></a></p><p id="par0110" class="elsevierStylePara elsevierViewall">The potential of certain pollutants to elicit allergic airway disease has been established in both experimental models and in humans&#46;<a class="elsevierStyleCrossRef" href="#bib0165"><span class="elsevierStyleSup">33</span></a> Short exposure &#40;10 days&#41; to HDM alone or to HDM and OVA resulted in equivalent levels of airway inflammation due to a robust inflammatory response in the lung&#46; Moreover&#44; it has been recently shown that intranasal administration of HDM daily for 10 days induces allergic sensitisation and Th2 airway inflammation&#44; both of which are substantially reduced by concurrent treatment with anti-M-CSF antibodies&#46;<a class="elsevierStyleCrossRef" href="#bib0170"><span class="elsevierStyleSup">34</span></a> This prompted us to expose the tested animals for about 10 days&#46;</p><p id="par0115" class="elsevierStylePara elsevierViewall">Allergic disorders have been rising in recent years&#46; It has been demonstrated that IgE antibodies play an important role in mediating type I hypersensitivity in humans&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">35</span></a> In both food and inhalant allergy it is accepted that food- or HDM-specific IgE binds to high-affinity Fc¿RI on mast cells&#44; basophiles&#44; macrophages&#44; and dendritic cells&#44; as well as to low-affinity Fc¿RII on macrophages&#44; monocytes&#44; lymphocytes&#44; eosinophils&#44; and platelets&#46;<a class="elsevierStyleCrossRef" href="#bib0180"><span class="elsevierStyleSup">36</span></a> When mite allergens penetrate mucosal barriers of the gastrointestinal or respiratory tract and contact IgE antibodies bound to mast cells or basophiles&#44; histamine and other mediators that induce symptoms of immediate hypersensitivity are released&#46;<a class="elsevierStyleCrossRef" href="#bib0185"><span class="elsevierStyleSup">37</span></a> Other studies have also suggested that cleavage of CD23 &#40;low-affinity IgE receptor&#41; may promote and enhance an IgE immune response&#46;<a class="elsevierStyleCrossRef" href="#bib0190"><span class="elsevierStyleSup">38</span></a></p><p id="par0120" class="elsevierStylePara elsevierViewall">Another factor to consider is cytokine production&#46; Indeed&#44; it has been suggested that HDM may privilege the generation of a Th2-polarised response by modulating the balance between IL-4 and IFN-&#947;&#46;<a class="elsevierStyleCrossRef" href="#bib0195"><span class="elsevierStyleSup">39</span></a> In addition&#44; various dust mite allergens act on bronchial epithelial cells in vitro to elicit the production of a number of cytokines&#46;<a class="elsevierStyleCrossRef" href="#bib0200"><span class="elsevierStyleSup">40</span></a></p><p id="par0125" class="elsevierStylePara elsevierViewall">IL-4 is a key anti-inflammatory cytokine and plays an important role in the cytokine network and in the Th1&#47;Th2 balance&#46; It is generated by helper T cells&#44; and possibly in the early adaptive immune response by mast cells and basophils&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">35</span></a> In this study&#44; the low level of IL-4 in the faeces-treated group may be due to the negative feedback that is exerted by histamine itself to inhibit IL-4&#46;<a class="elsevierStyleCrossRef" href="#bib0185"><span class="elsevierStyleSup">37</span></a></p><p id="par0130" class="elsevierStylePara elsevierViewall">Thus&#44; this study tended to measure the levels of INF-&#947;&#44; L-4 and IgE that are involved in allergic disease&#46; Repeated exposure to <span class="elsevierStyleItalic">T&#46; putrescentiae</span> allergic faeces caused an increase in both INF-&#947; and IgE&#44; and a decrease in IL-4&#46; The high level of both INF-&#947; and IgE&#44; and the low level of IL-4 may participate in airway disorder&#46; After garlic feeding&#44; the increase of IL-4 and the decrease of both INF-&#947; and IgE were maintained&#44; and this may play an important role in the airway remodelling processes&#46; The immunomodulatory effects on INF-&#947;&#44; IL-4 and IgE balance were tested by others&#46;<a class="elsevierStyleCrossRefs" href="#bib0070"><span class="elsevierStyleSup">14&#44;41</span></a></p><p id="par0135" class="elsevierStylePara elsevierViewall">Garlic in small amounts applied as a food condiment does not usually pose an important risk&#44; except for rare specific allergic reactions in hypersensitive people&#46;<a class="elsevierStyleCrossRef" href="#bib0210"><span class="elsevierStyleSup">42</span></a> In contrast&#44; this study showed that some allergic disorder could be modulated by garlic&#46;</p><p id="par0140" class="elsevierStylePara elsevierViewall">In conclusion&#44; these data demonstrated that wheat germ&#44; roomy cheese&#44; nido&#44; egg yolk and potatoes media increase the mass rearing of <span class="elsevierStyleItalic">T&#46; putrescentiae</span> compared to those recorded by fish meal&#44; copra&#44; egg albumin&#44; chicken stocks&#44; pollens&#44; bean&#44; human hair &#38; scales and yeast extract&#46; Repeated exposure to <span class="elsevierStyleItalic">T&#46; putrescentiae</span> allergic faeces caused an increase in both the pro-inflammatory cytokine INF-&#947; and the main allergic immunoglobulin IgE&#44; and a decrease in the anti-inflammatory cytokine IL-4&#46; The high level of both INF-&#947; and IgE&#44; and the low level of IL-4 may participate in airway disorder&#46; After garlic treating&#44; the increase of IL-4 and the decrease of both INF-&#947; and IgE were maintained&#44; and this means that garlic may play an important role in the amelioration of the airway immune response&#46;</p></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Ethical disclosures</span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Protection of human and animal subjects</span><p id="par0145" class="elsevierStylePara elsevierViewall">The procedures followed were in accordance with the regulations of the responsible Clinical Research Ethics Committee and in accordance with those of the World Medical Association and the Helsinki Declaration&#46;</p></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Confidentiality of data</span><p id="par0150" class="elsevierStylePara elsevierViewall">No patient data appears in this article&#46;</p></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Right to privacy and informed consent</span><p id="par0155" class="elsevierStylePara elsevierViewall">No patient data appears in this article&#46;</p></span></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Conflict of interest</span><p id="par0160" class="elsevierStylePara elsevierViewall">The author has no conflict of interest to declare&#46;</p></span></span>"
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          "titulo" => "Introduction"
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          "titulo" => "Materials and methods"
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              "identificador" => "sec0015"
              "titulo" => "Mass rearing of Tyrophagus putrescentiae"
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              "titulo" => "Crude extracts"
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              "titulo" => "Experimental animals"
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          "titulo" => "Results"
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              "identificador" => "sec0040"
              "titulo" => "Mass rearing of Tyrophagus putrescentiae"
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              "titulo" => "Quantification of INF-&#947;"
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              "titulo" => "Quantification of IL-4"
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              "titulo" => "Quantification of IgE"
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          "titulo" => "Discussion"
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          "titulo" => "Ethical disclosures"
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              "titulo" => "Protection of human and animal subjects"
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              "titulo" => "Confidentiality of data"
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            2 => array:2 [
              "identificador" => "sec0080"
              "titulo" => "Right to privacy and informed consent"
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          "titulo" => "Conflict of interest"
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          "titulo" => "Acknowledgements"
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          "titulo" => "References"
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    "fechaRecibido" => "2013-03-09"
    "fechaAceptado" => "2013-05-10"
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            0 => "Mass rearing"
            1 => "Natural diet"
            2 => "<span class="elsevierStyleItalic">T&#46; putrescentiae</span> faeces"
            3 => "Allergic immune response"
            4 => "IgE"
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        "titulo" => "Abstract"
        "resumen" => "<span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">The storage mite&#44; <span class="elsevierStyleItalic">Tyrophagus putrescentiae</span>&#44; detected in the samples collected from stored products and house dust&#44; is one of the major causes of allergic disorders&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0015">Objective</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">The purpose of this study was to ameliorate the <span class="elsevierStyleItalic">T&#46; putrescentiae</span> faeces allergic immunological disorder by garlic&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Albino experimental rats were classified into control&#44; inhaled and treated groups&#46; Mass rearing of <span class="elsevierStyleItalic">T&#46; putrescentiae</span> on different diets&#44; and ELISA of some cytokines and IgE techniques were used&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">The results obtained showed the highest population of <span class="elsevierStyleItalic">T&#46; putrescentiae</span> reared in four from thirteen tested diets&#46; In addition&#44; significantly higher serum levels of INF-&#947; and IgE were found in rats treated with faeces than the other groups&#59; especially the garlic-treated group&#46; In contrast&#44; IL-4 was lower in faeces-treated rats than the others&#59; however&#44; the control group had the highest level of IL-4&#46; Statistical analysis of data showed a significant difference between the garlic-treated group and either control or faeces-treated groups &#40;<span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0030">Conclusions</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">The population of <span class="elsevierStyleItalic">T&#46; putrescentiae</span> mites peaked in four from thirteen tested diets&#46; The immunological disorder caused by repeated exposure to <span class="elsevierStyleItalic">T&#46; putrescentiae</span> faeces might be modulated by garlic&#46;</p>"
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Article information

ISSN: 03010546
Original language: English

DOI:10.1016/j.aller.2013.05.002

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Year/Month	Html	Pdf	Total

2024 November	10	0	10
2024 October	61	6	67
2024 September	107	3	110
2024 August	84	11	95
2024 July	81	8	89
2024 June	78	1	79
2024 May	90	14	104
2024 April	71	8	79
2024 March	90	13	103
2024 February	104	8	112
2024 January	143	10	153
2023 December	178	3	181
2023 November	204	14	218
2023 October	184	13	197
2023 September	104	9	113
2023 August	82	8	90
2023 July	85	26	111
2023 June	73	6	79
2023 May	74	9	83
2023 April	81	6	87
2023 March	65	4	69
2023 February	67	7	74
2023 January	72	35	107
2022 December	66	11	77
2022 November	70	9	79
2022 October	43	13	56
2022 September	39	9	48
2022 August	56	18	74
2022 July	46	7	53
2022 June	56	14	70
2022 May	60	7	67
2022 April	49	14	63
2022 March	71	29	100
2022 February	40	8	48
2022 January	94	11	105
2021 December	54	16	70
2021 November	46	14	60
2021 October	46	22	68
2021 September	73	19	92
2021 August	94	10	104
2021 July	79	9	88
2021 June	51	19	70
2021 May	57	9	66
2021 April	73	14	87
2021 March	28	22	50
2021 February	42	20	62
2021 January	43	13	56
2020 December	1	2	3
2020 August	0	1	1
2020 July	0	1	1
2020 May	1	4	5
2020 March	0	1	1
2020 February	0	1	1
2020 January	0	3	3
2019 October	0	2	2
2019 September	0	4	4
2019 August	0	3	3
2019 July	0	13	13
2019 June	0	6	6
2019 May	0	17	17
2018 February	17	2	19
2018 January	17	2	19
2017 December	9	1	10
2017 November	16	5	21
2017 October	25	6	31
2017 September	14	11	25
2017 August	18	6	24
2017 July	20	12	32
2017 June	62	10	72
2017 May	33	8	41
2017 April	18	4	22
2017 March	21	43	64
2017 February	20	2	22
2017 January	23	5	28
2016 December	22	3	25
2016 November	19	4	23
2016 October	30	7	37
2016 September	30	12	42
2016 August	23	6	29
2016 July	17	3	20
2016 June	27	10	37
2016 May	22	11	33
2016 April	25	27	52
2016 March	14	6	20
2016 February	12	10	22
2016 January	13	10	23
2015 December	14	6	20
2015 November	15	5	20
2015 October	16	4	20
2015 September	11	5	16
2015 May	0	1	1
2015 February	1	2	3
2014 November	2	1	3
2014 September	1	1	2

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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