Effect of host genotype and Eimeria acervulina infection on the metabolome of meat-type chickens.
Aggrey Samuel E,Milfort Marie C,Fuller Alberta L,Yuan Jianmin,Rekaya Romdhane
OBJECTIVE:A study was conducted to identify metabolic biochemical differences between two chicken genotypes infected with Eimeria acervulina and to ascertain the underlying mechanisms for these metabolic alterations and to further delineate genotype-specific effects during merozoite formation and oocyst shedding. METHODS:Fourteen day old chicks of an unimproved (ACRB) and improved (COBB) genotype were orally infected with 2.5 x 105 sporulated E. acervulina oocysts. At 4 and 6 day-post infection, 5 birds from each treatment group and their controls were bled for serum. Global metabolomic profiles were assessed using ultra performance liquid chromatography/tandem mass spectrometry (metabolon, Inc.,). Statistical analyses were based on analysis of variance to identify which biochemicals differed significantly between experimental groups. Pathway enrichment analysis was conducted to identify significant pathways associated with response to E. acervulina infection. RESULTS:A total of 752 metabolites were identified across genotype, treatment and time post infection. Altered fatty acid (FA) metabolism and β-oxidation were identified as dominant metabolic signatures associated with E. acervulina infection. Key metabolite changes in FA metabolism included stearoylcarnitine, palmitoylcarnitine and linoleoylcarnitine. The infection induced changes in nucleotide metabolism and elicited inflammatory reaction as evidenced by changes in thromboxane B2, 12-HHTrE and itaconate. CONCLUSIONS:Serum metabolome of two chicken genotypes infected with E. acervulina demonstrated significant changes that were treatment-, time post-infection- and genotype-dependent. Distinct metabolic signatures were identified in fatty acid, nucleotide, inflammation and oxidative stress biochemicals. Significant microbial associated product alterations are likely to be associated with malabsorption of nutrients during infection.
Anti-Coccidial Effect of Rumex Nervosus Leaf Powder on Broiler Chickens Infected with Eimeria Tenella Oocyst.
Qaid Mohammed M,Al-Mufarrej Saud I,Azzam Mahmoud M,Al-Garadi Maged A,Albaadani Hani H,Alhidary Ibrahim A,Aljumaah Riyadh S
Animals : an open access journal from MDPI
Coccidiosis a huge economic burden in poultry farms where the pathogen harms animal well-being and survival. Besides synthetic anti-coccidial drugs, natural herbs appear to be an alternative way to prevent avian coccidiosis. (RN), a phytogenic shrub, has received considerable attention in recent years due to its significant anti-microbial effects; however, limited knowledge exists about its potential anti-coccidial functions. This study was conducted to evaluate the prophylactic and therapeutic activities of RN leaf powder in broilers infected with . Infected chickens received a commercial diet containing 1, 3, or 5 g RN powder/kg diet compared to infected broilers that treated with Sacox (PC) or compared to uninfected broilers that received a commercial diet alone (NC). Results showed that RN powder significantly ( < 0.05) reduced the lesion scores and suppressed the output of oocysts per gram (OPG) in chickens' feces. Although RN was unable to minimize the weight gain loss due to emeriosis, RN at level 1 g improved the feed conversion ratio. Therefore, RN powder, at 5 g, possesses moderate anti-coccidial effects and hence could be used to treat avian coccidiosis in field conditions; however, further studies are required to investigate, in vitro or in vivo, the anti-coccidial potential of active ingredients.
Anticoccidial effectivity of a traditional medicinal plant, Cinnamomum verum, in broiler chickens infected with Eimeria tenella.
Qaid Mohammed M,Al-Mufarrej Saud I,Azzam Mahmoud M,Al-Garadi Maged A
The anticoccidial influences of various amounts of Cinnamomum verum powder were compared with that of salinomycin as standard synthetic anticoccidial drug on the anticoccidial indicators and production performance in broilers experimentally exposed to coccidiosis. Broiler chicks at 1 d of age (n = 150) were arbitrarily distributed into 6 groups. Birds from groups 1-3 were received the starter and finisher diets plus 2, 4, and 6 g of cinnamon/kg of the diet, respectively. Birds from group 4 were fed the starter and finisher diets plus 66 mg of salinomycin, group 5 constituted the positive control (PC), with the coccidial challenge, and group 6 constituted the negative control (NC), without the coccidial infection, which were both maintained on diets without any cinnamon. The results showed that BW gain, feed conversion ratio, and production efficiency factor declined considerably (P < 0.05) in the PC compared with the NC. At seventh day postinfection (DPI), the lesion score was significantly (P < 0.05) lower in the ceca of salinomycin-treated birds than the PC. The anticoccidial index was moderate in the chickens treated with salinomycin and 6 g cinnamon at 7 DPI compared with those in the PC group. In addition, cinnamon- or salinomycin-treated birds exhibited lower oocyst values and higher oocyst reduction rate than those in the PC. We concluded that C. verum at level 6 g cinnamon/kg diet moderately reduced coccidiosis and attempted to improved BW, feed conversion ratio, and production efficiency at 7 DPI compared with the infected groups.
Anticoccidial effect of extract against .
Yong Ting,Chen Meng,Li Yunhe,Song Xu,Huang Yongyuan,Chen Yaqin,Jia Renyong,Zou Yuanfeng,Li Lixia,Yin Lizi,He Changliang,Lv Cheng,Liang Xiaoxia,Ye Gang,Yin Zhongqiong
CONTEXT: extracts (FMTE) have a good therapeutic effect on coccidiosis, but there is no relevant research on its prophylactic effect on coccidiosis. OBJECTIVE:This study comprehensively evaluates the anticoccidial effect of FMTE. MATERIALS AND METHODS:, the unsporulated oocysts were treated with serial dilutions of FMTE and incubated for 7 d, and the sporulated oocysts were counted for calculating the median lethal concentration (LC) of FMTE. , 180 10-day-old broiler chickens free of coccidiosis were weighted and randomly distributed into six groups: normal group, untreated group, 4 protective groups (positive group and three FMTE groups). From day 10 to day 21, chickens in the three FMTE groups were pre-treated with FMTE at the dosage of 2.5, 5 and 10 g/kg/d, respectively, and chickens in the positive group were pre-treated with qiuliling (10 g/kg/d). On day 14, chickens in all groups except the normal group were orally infected with 1.5 × 10 sporulated oocysts. The clinical symptoms were observed from day 10 to day 21, the anticoccidial index (ACI), tissue lesions, and intestinal microflora were determined on day 21. RESULTS:FMTE showed anti-sporulation effect against and the LC value was 245.83 µg/mL . , FMTE at the dosage of 10 g/kg/d was effective against infection, and its ACI value was 162.56, which was higher than the value of positive drug qiuliling (128.81). FMTE have potent anticoccidial effects, and it presents an alternative anticoccidial agent for avian coccidiosis control.
Nucleotide-rich yeast extract fed to broiler chickens challenged with Eimeria: impact on growth performance, jejunal histomorphology, immune system, and apparent retention of dietary components and caloric efficiency1.
Leung H,Patterson R,Barta J R,Karrow N,Kiarie E
Nucleotide-rich yeast extract (YN) was investigated for its effects on growth performance, jejunal histomorphology and mucosal immunoglobulin A (IgA), immune organs weight and apparent retention (AR) of components in broiler chickens challenged with Eimeria. A total of 336 day-old male chicks (Ross x Ross 708) were placed in floor pens and provided a corn-soybean meal-based diet without or with YN (500 g/mt) (n = 14). On day 10, 7 replicates per diet were orally administered with 1 mL of sporulated E. acervulina and E. maxima oocysts and the rest (non-challenged control) administered equivalent distilled water creating a 2 × 2 factorial arrangement for the post-challenge period (day 11 to 35). Feed intake (FI), BWG, and FCR responses were measured in pre- and post-challenge periods. Excreta samples were collected on day 14 to 17 and 31 to 34 for oocyst count and AR of components, respectively. On day 15 and 35, 5 birds/pen were necropsied for intestinal samples. Spleen, bursa, and thymus weights were also recorded at both time points and breast yield on day 35. Diet had no effect (P > 0.05) on pre-challenge growth performance. Interaction (P = 0.046) between Eimeria and YN on FI was such that Eimeria challenge increased FI (day 11 to 35) in non-YN birds. There was no interaction (P > 0.05) between Eimeria and YN on other post-challenge responses. Eimeria reduced (P < 0.05) BWG, FCR, caloric efficiency, day 15 jejunal villi height and IgA concentration, and increased (P < 0.01) day 15 spleen weight. On day 35, YN increased bursa weight (1.57 vs. 1.78 mg/g BW, P = 0.04). There was a tendency for an interaction effect (P = 0.09) on day 35 thymus weight, such that in challenged birds, YN fed birds tended to have a lighter thymus relative to non-YN fed birds. In conclusions, independent of Eimeria challenge, supplemental YN had no effect on growth performance, caloric efficiency, and intestinal function but increased immune organ weights.
Pleurotus ostreatus extract inhibits Eimeria species development in naturally infected broiler chickens.
Ademola Isaiah O,Ojo Philip O,Odeniran Paul O
Tropical animal health and production
Coccidian organisms of poultry have proved very hard to control due to their abilities to resist most anticoccidial drugs. Pleurotus ostreatus (Fr.) Jacq. ex (Pleurotaceae), a medicinal mushroom, was investigated in vivo against Eimeria spp. Ninety-six broilers (day-old) naturally infected with Eimeria spp. were divided into eight groups (12 birds per group). Group A was infected untreated (negative control) and group B was treated with toltrazuril (positive control) while groups C-H were gavaged with graded doses of P. ostreatus extract at 100, 200, 300, 400, 500, and 600 mg/kg, respectively. The phytochemical analysis of the aqueous extract of P. ostreatus which revealed saponins, flavonoids, anthraquinones, and alkaloids was evaluated for anticoccidial activity by assessing the inhibition of oocyst output, lesion score, faecal score, weight differences, haematological parameters, and leucocyte differential counts. The acute toxicity study showed extract of P. ostreatus to be non-toxic at 600 mg/kg. The weight of the groups treated with the extracts and toltrazuril increased significantly (P < 0.05) compared with the untreated control. Treated groups significantly (P < 0.05) reduced oocyst output except groups C and D. The therapeutic best-fit ED value for the extract was 448 mg/kg. The post-treatment mean packed cell volume and red blood cell count were significantly higher (P < 0.05) than the untreated group, while the WBC count was significantly higher (P < 0.05) in the untreated group. Pleurotus ostreatus therefore could be a potential source of new anticoccidial medicine which could find application in the control of avian coccidiosis.
Effects of tannic acid extract on performance and intestinal health of broiler chickens following coccidiosis vaccination and/or a mixed-species Eimeria challenge.
Tonda R M,Rubach J K,Lumpkins B S,Mathis G F,Poss M J
Two experiments were conducted to investigate the effects of tannic acid extract (TAE) formulations on the performance and intestinal health of male Cobb × Cobb 500 broilers exposed to coccidiosis. In the first experiment, 320 broiler chicks were randomly assigned to 5 treatments with 8 replicates. Treatments included non-medicated, uninfected (NC); non-medicated, infected (PC); salinomycin (SAL, 66 mg/kg); tannic acid (TA, 0.5 g/kg) and TAE (TAE, 0.5 g/kg). On d 14, all groups (except NC) were orally inoculated with Eimeria acervulina, E. maxima and E. tenella oocysts. Intestinal lesion scores, fecal oocyst counts (OPG) and performance were evaluated on d 20. The PC had greater lesions and higher FCR than infected, supplemented groups. Only TAE reduced OPG compared to PC (P < 0.05). In the second experiment, 3,000 broiler chicks were vaccinated on day of hatch with live coccidial oocysts, then randomly assigned to 5 treatments with 15 replicates. Treatments included non-medicated (CNT); salinomycin (SAL, 66 mg/kg); robenidine (ROB, 33 mg/kg); TAE (0.5 g/kg) and TAE with Bacillus coagulans (TAE+BC, 0.5 g/kg). On d 29, a subset of pens (n = 20) were challenged with a mixed Eimeria spp. oral inoculum; performance, lesions and OPG were evaluated on d 35. An immune challenge was created in half the pens by issuing broilers feed without supplementation materials during the challenge. For the non-challenged pens (n = 55), performance was measured up to d 49. Performance of non-challenged, vaccinated-CNT birds was improved with all treatments at d 21 and d 49. Among the challenged birds, withdrawal of SAL or ROB resulted in FCR similar to the challenged CNT group (P > 0.05), whereas withdrawal of TAE or TAE+BC maintained improved FCR compared to challenged-CNT birds (P < 0.05). These findings indicate supplementation of TAE and TAE+BC with coccidiosis vaccination can be considered as a potential alternative strategy to address coccidiosis in broiler chickens.
Antioxidant and anti-inflammatory activities of pomegranate (Punica granatum) on Eimeria papillata-induced infection in mice.
Amer Omar S O,Dkhil Mohamed A,Hikal Wafaa M,Al-Quraishy Saleh
BioMed research international
Coccidiosis is the most prevalent disease causing widespread economic loss, especially in poultry farms. Here, we investigated the effects of pomegranate peel extract (PPE) on the outcome of coccidiosis caused by Eimeria papillata in mice. The data showed that mice infected with E. papillata and treated with PPE revealed a significant decrease in the output of oocysts in their faeces by day 5 p.i. Infection also induced inflammation and injury of the jejunum. This was evidenced (i) as increases in reactive oxygen species, (ii), as increased neutrophils and decreased lymphocytes in blood (ii) as increased mRNA levels of inducible nitric oxide synthase (iNOS), Bcl-2 gene, and of the cytokines interferon gamma (IFN-γ), tumour necrosis factor-α (TNF-α), and interleukin-1β (IL-1β), and (iv) as downregulation of mucin gene MUC2 mRNA. All these infection-induced parameters were significantly altered during PPE treatment. In particular, PPE counteracted the E. papillata-induced loss of the total antioxidant capacity. Our data indicated that PPE treatment significantly attenuated inflammation and injury of the jejunum induced by E. papillata infections.
Anticoccidial efficacy of Canary rue (Ruta pinnata) extracts against the caprine apicomplexan Eimeria ninakohlyakimovae.
López Adassa M,Muñoz María C,Molina José M,Hermosilla Carlos,Taubert Anja,Zárate Rafael,Hildebrandt Inna,McNaughton-Smith Grant,Eiroa José L,Ruiz Antonio
Journal of animal science
Continuous use of anticoccidial treatments against Eimeria infections has resulted in the development of drug resistance. This study aimed to evaluate the anticoccidial efficacy of a methanolic extract derived from the endemic Canary rue (Ruta pinnata) plant of the Canary Islands, Spain, against Eimeria ninakohlyakimovae using in vitro assays. Freshly unsporulated oocysts were exposed to different concentrations of R. pinnata extract and thereafter evaluated for sporulation inhibition. Additionally, anticoccidial activity was examined by testing the viability of the E. ninakohlyakimovae sporozoites and their ability to infect bovine colonic epithelial cells after incubation with different concentrations of R. pinnata plant extract. The inhibition of oocyst sporulation by the extract was both time and concentration dependent, with certain combinations affording the same levels of sporulation inhibition as formaldehyde used as positive control (P < 0.001). Moreover, concentrations >0.1 mg/mL also affected not only the viability of the sporozoites but also their cell invasion capacity (P < 0.001). Altogether, these results show that methanolic fruit extracts from R. pinnata have important anticoccidial activity against oocysts and sporozoites of Eimeria. The potential efficacy of the extracts against other animal/human parasites remains to be elucidated, and further studies are needed to better understand its mode of action against coccidian parasites.
Anticoccidial effects of Aloe secundiflora leaf extract against Eimeria tenella in broiler chicken.
Kaingu Fredrick,Liu Dandan,Wang Lele,Tao Jianping,Waihenya Rebecca,Kutima Helen
Tropical animal health and production
Anticoccidial effects of Aloe secundiflora crude leaf extract was tested in broiler chickens following oral infection with Eimeria tenella. Sixty 22-day-old birds were divided into six groups of ten birds each. Three treatment groups A, B, and C were fed with the extract (100, 250, and 500 mg/day, respectively) mixed in feed for 10 days, and three control groups: group D (drug control) administered 300 mg/l of sulfachloropyrazine sodium soluble powder in drinking water for 5 days, group E (infected/non-medicated positive control), and group F (uninfected/non-medicated negative control). Except for group F, all groups were orally inoculated with 75,000 sporulated oocysts of E. tenella. The effects of the extract on E. tenella infection were evaluated by severity of bloody diarrhea, body weight (BW) gain, oocyst output, and lesion score. No bird in the treated groups died of coccidiosis, and severity of bloody diarrhea was milder than in the positive control group. BW gains in the treated groups were significantly higher than in group E (p < 0.05). The lesion scores of the treated groups were significantly lower than that of group E. Oocyst output in groups A, B, and C were 11.23, 8.24, and 6.82 × 10, respectively. As compared with the negative control group (12.84 × 10), the reductions in oocyst production were 12.54, 35.83, and 46.88%, respectively. Oocyst output significantly reduced with an increase in Aloe dosage. The findings of this study suggest that Aloe secundiflora extract presents an alternative anticoccidial agent for the control of avian coccidiosis.
Effects of probiotic and Bidens pilosa on the performance and gut health of chicken during induced Eimeria tenella infection.
Memon F U,Yang Y,Lv F,Soliman A M,Chen Y,Sun J,Wang Y,Zhang G,Li Z,Xu B,Gadahi J A,Si H
Journal of applied microbiology
AIM:In this study, we have examined the individual and combined protective mechanism of probiotic and Bidens pilosa on the performance and gut health of chickens during Eimeria tenella infection over a 29-day experimental trial. METHODS AND RESULTS:A total of one hundred and fifty 1-day-old chickens were equally distributed into five treatment groups with three biological replicates: two groups were allocated as control groups (control group untreated unchallenged, CG and control positive untreated challenged, CPG) and three groups were fed diets with probiotic (PG), B. pilosa (BPG) and probiotic + B. pilosa (PG + BPG) and challenged with E. tenella. Birds of all groups were assessed for pre and post-infection body weights, oocysts shedding, caecal lesion scores and mRNA expression levels of apoptosis related proteins (Bcl-2, Bax and caspase-3), antioxidant enzymes (CAT and SOD 1), pro-inflammatory cytokines (IL-6 and IL-8) and tight junction proteins (CLDN 1 and ZO 1). Our results revealed that during infection (day 21-29), E. tenella challenged chickens significantly decreased the body weight compared with uninfected control chickens; however, there was no significant effect on body weight of chickens fed with probiotic, B. pilosa and probiotic + B. pilosa was observed. Eimeria tenella challenged untreated birds increased (P < 0·05) oocysts shedding, destructive ratio of caeca and mortality as compared to treated challenged birds. CPG group up-regulated the mRNA expression levels of anti-apoptosis protein Bcl-2 while down-regulated the pro-apoptosis protein Bax relative to PG, BPG and PG + BPG groups. Moreover chickens fed probiotic, B. pilosa and probiotic + B. pilosa diets enhanced the activities of antioxidant enzymes, pro-inflammatory cytokines and tight junction proteins with the comparison of control positive untreated challenged chickens. CONCLUSION:These findings elaborated that feed supplementation of probiotic and B. pilosa (individually or in combination) appeared to be effective in inhibiting the occurrence of disease and decreasing the severity of Eimeria infection in chickens. SIGNIFICANCE AND IMPACT OF THE STUDY:This study explained the underlying anti-coccidial mechanism in which probiotic and B. pilosa (individually and/or in combination) improve the performance of chicken and protect against gut inflammatory responses caused by E. tenella.
Immunological evaluation of invasive Lactobacillus plantarum co-expressing EtMIC2 and chicken interleukin-18 against Eimeria tenella.
Zhang Zan,Huang Hai-Bin,Jiang Yan-Long,Liu Jing,Gao Xing,Liu Yang,Yang Wen-Tao,Shi Chun-Wei,Wang Dan,Wang Jian-Zhong,Kang Yuan-Huan,Wang Chun-Feng,Yang Gui-Lian
Chicken coccidiosis is a protozoan parasitic disease that leads to considerable economic losses in the poultry industry. In this study, we used invasive Lactobacillus plantarum (L.P) expressing the FnBPA protein as a novel bacterial carrier for DNA delivery into epithelial cells to develop a live oral DNA vaccine. A fusion DNA vaccine co-expressing EtMIC2 and chicken IL-18 (chIL-18) was constructed and then delivered to the host by invasive L.P. Its efficacy against Eimeria tenella challenge was evaluated in chickens by examining the relative weight gain rate; caecal lesion score; OPG; anti-coccidial index (ACI); levels of EtMIC2 antibody, FnBPA, IL-4, IL-18, IFN-γ and SIgA; and proliferation ability and percentages of CD4 and CD8 splenocytes. The experimental results showed that chickens immunized with invasive L.P carrying the eukaryotic expression vector pValac-EtMIC2 (pValac-EtMIC2/pSIP409-FnBPA) had markedly improved immune protection against challenge compared with that of chickens immunized with non-invasive L.P (pValac-EtMIC2/pSIP409). However, invasive L.P co-expressing EtMIC2 with the chIL-18 vector exhibited the highest protection efficiency against E. tenella. These results indicate that invasive Lactobacillus-expressing FnBPA improved humoural and cellular immunity and enhanced resistance to E. tenella. The DNA vaccine delivered by invasive Lactobacillus provides a new concept and method for the prevention of E. tenella.
Effect of different floatation solutions on E. tenella oocyst purification and optimization of centrifugation conditions for improved recovery of oocysts and sporocysts.
Qi Nanshan,Liao Shenquan,Abuzeid Asmaa M I,Li Juan,Wu Caiyan,Lv Minna,Lin Xuhui,Hu Junjing,Xiao Wenwan,Sun Mingfei,Li Guoqing
Saturated salt floatation method is widely used for coccidian oocyst purification. However, the repeated procedures and inefficient oocysts recovery rate are a continuous challenge. This study aimed to investigate the best suitable floatation solution, along with optimal centrifugation speed and time for Eimeria tenella (E. tenella) oocyst and sporocyst purification. Different floatation solutions i-e, saturated salt, Sheather's sugar and sodium hypochlorite (NaClO) at 20-60% concentrations were used to purify oocyst. It was found that about 96.99% oocysts (8609×g for 10 min) were recovered under these conditions without any effect on the viability of sporocysts. The recovery rate of oocysts using 50% NaClO (V/V) was significantly higher than 35% saturated salt flotation solution (P < 0.05). The optimal method for purification of oocysts based our experimentation was centrifugation at 8609×g for 3 min using 50% NaClO floatation solution, and the optimized centrifugation conditions for improved recovery of sporocysts (about 99.3%) were at 2152×g for 5 min. The present study provided a better method for the coccidian oocyst purification, which could be successfully adopted as a better alternative to existing techniques commonly used for investigations/research pertaining to coccidia.
Eimeria tenella infection perturbs the chicken gut microbiota from the onset of oocyst shedding.
Huang Guangping,Tang Xiaoli,Bi Feifei,Hao Zhenkai,Han Zhenyan,Suo Jingxia,Zhang Sixin,Wang Si,Duan Chunhui,Yu Zhengquan,Yu Fang,Yu Yonglan,Lv Yanli,Suo Xun,Liu Xianyong
Coccidiosis is a serious threat to the poultry industry, resulting in substantial economic losses worldwide. The effective development of alternative treatments for coccidiosis that does not involve chemotherapy drugs and does not result in antibiotic resistance relies on gaining a clearer understanding of the interaction between host intestinal microbiota and enteric coccidia. Here, we established an Eimeria tenella infection model in chickens and subsequently monitored the changes in the overall intestinal microbiome using 16S rRNA gene sequencing. We found that the gut (i.e. fecal) microbiota of infected chicken differed from that of uninfected naïve animals. Levels of non-pathogenic bacteria, including Lactobacillus and Faecalibacterium declined, whereas those of pathogenic bacteria, including Clostridium, Lysinibacillus, and Escherichia, increased over time in response to E. tenella infection. Similar dynamic changes of the fecal microbiota were observed in both Arbor Acres broilers and White Leghorn chickens, indicating that the perturbation of the microbiota was directly induced by E. tenella infection. Our findings could be used to further elucidate the serious damage to host health caused by coccidia infection, leading to the development of new effective treatment options for coccidiosis.
In vitro efficacy of allicin on chicken Eimeria tenella sporozoites.
Alnassan Alaa Aldin,Thabet Ahmed,Daugschies Arwid,Bangoura Berit
Chicken coccidiosis is a major parasitic disease caused by Eimeria spp. It is controlled and treated using chemical anticoccidial agents. Development of partial or complete resistance toward these anticoccidials is considered a major problem in poultry industry. Allicin is an organosulfur compound produced as a result of the reaction between alliin and alliinase after hacking of garlic. In this study, tenfold dilution from 180 mg/ml to 1.8 ng/ml of allicin in distilled water was tested against E. tenella in vitro. The percent of inhibition in allicin was from 99.9 to 71.53% using 180 mg/ml and 180 ng/ml, respectively. The percent of inhibition was 56.24% using 1.8 ng/ml. We used allicin as a treatment from plants against chicken coccidiosis; however, in vivo study should be performed to confirm these results.
Anticoccidial effects of herbal extracts on Eimeria tenella infection in broiler chickens: in vitro and in vivo study.
Habibi Hasan,Firouzi Sobhan,Nili Hasan,Razavi Mostafa,Asadi Seyedeh Leili,Daneshi Sajad
Journal of parasitic diseases : official organ of the Indian Society for Parasitology
Safe alternative anticoccidial drug to chemical feed additives are herbal extracts, because they don't results to tissue residue and drug resistance. In order to evaluate the effects of herbal extracts to control avian coccidiosis, 180 one-day-old broiler chickens were randomly divided into nine equal groups, as follows: (1) Biarum bovei (2) Nectaroscordum tripedale( 3) Dorema aucheri (4) Cichorium intybus (5) Prangos ferulaceae (6) diclazuril (7) Artemisia absinthium (8) infected control (9) uninfected control (each contains two groups). Administration of herbal extracts and supplementation of diclazuril was began 2 days before challenge and lasted for the duration of the experiment. The chicks of all the groups except uninfected control group were inoculated orally with sporulated oocysts (3 × 10(3) oocysts of Eimeria tenella) on the day 22 of age. The criteria employed were: body weight, feed conversion ratio, blood in feces, survival rate, lesion scoring, number of oocyst output per gram feces and histopathological changes. For histopathological evaluation, on day 12 post inoculation three birds from each group were randomly selected and humanly sacrificed. N. tripedale and diclazuril revealed better results in terms of growth performance, lesion score, extent of bloody diarrhea and oocyst count as compared to other herbal extracts. The increase in the severity of lesions was observed in groups of D. aucheri, A. absinthium, B. bovei, P. ferulaceae, C. intybus, diclazuril and N. tripedale, respectively. In conclusion, the current study showed that herbal extracts were effective in control of coccidiosis caused by the E. tenella infection.
Anticoccidial activity of herbal complex in broiler chickens challenged with Eimeria tenella.
Zaman Muhammad A,Iqbal Zafar,Abbas Rao Z,Khan Muhammad N
The objective of the present study was to evaluate the anticoccidial effect of different concentrations of the herbal complex of 4 plants (leaves of Azadirachta indica and Nicotiana tabacum, flowers of Calotropis procera and seeds of Trachyspermum ammi) in broiler chickens in comparison with amprolium anticoccidial. Three concentrations (2 g, 4 g and 6 g) of herbal complex were given to the experimental groups once a day and amprolium (at the dose rate of 125 ppm) was given orally in drinking water from the 14th to the 21st days of age. One group was kept as infected, non-medicated control and one as non-infected, non-medicated control. All groups were inoculated orally with 75,000 sporulated oocysts on the 14th day of age except the non-infected, non-medicated control. Among herbal complex medicated groups, the maximum anticoccidial effect was seen in the group medicated with 6 g herbal complex followed by 4 g and 2 g herbal complex medicated groups. Treatment with 6 g of the herbal complex significantly reduced the negative performance and pathogenic effects associated with Eimeria tenella challenge at a level that was comparable with amprolium when using a largely susceptible recent field isolate. In summary, concentration-dependent anticoccidial activity of the studied herbal complex suggests its use as an alternative anticoccidial agent to chemotherapeutic drugs for Eimeria tenella control.
Anticoccidial activity of traditional Chinese herbal Dichroa febrifuga Lour. extract against Eimeria tenella infection in chickens.
Zhang De-Fu,Sun Bing-Bing,Yue Ying-Ying,Zhou Qian-Jin,Du Ai-Fang
The study was conducted on broiler birds to evaluate the anticoccidial efficacy of an extract of Chinese traditional herb Dichroa febrifuga Lour. One hundred broiler birds were assigned to five equal groups. All birds in groups 1-4 were orally infected with 1.5 × 10(4) Eimeira tenella sporulated oocysts and birds in groups 1, 2 and 3 were medicated with 20, 40 mg extract/kg feed and 2 mg diclazuril/kg feed, respectively. The bloody diarrhea, oocyst counts, intestinal lesion scores, and the body weight were recorded to evaluate the anticoccidial efficacy. The results showed that D. febrifuga extract was effective against Eimeria infection; especially 20 mg D. febrifuga extract/kg feed can significantly increase body weight gains and reduce bloody diarrhea, lesion score, and oocyst excretion in comparison to infected-unmedicated control group.
Effects of a herbal complex against Eimeria tenella infection in chickens.
Du A,Hu S
Journal of veterinary medicine. B, Infectious diseases and veterinary public health
A liquid and a powder made from a herbal complex consisting of Uncariae Ramulus cum Uncis, Agrimoniae Herba, Sanguisorbae Radix, Eclipta Prostrate Herba, Pulsatillae Radix, Sophorae Flavescentis Radix, Rehmanniae Radix and Glycyrrhizae Radix were studied for their anticoccidial activities in chickens. Chickens were administered with herbal liquid, powder, diclazuril or without medication during the study and challenged with oocysts of Eimeria tenella. Results indicated that the birds medicated showed less bloody faeces than those without medication. The intestinal lesion was mild in the chicks medicated with herbal liquid without significantly different lesion score when compared with uninfected chicks. The birds with medication had significantly higher body weight gains than birds without medication. Therefore, the herbal complex used in this study was effective against E. tenella infection in chickens.
The effects of different doses of curcumin compound on growth performance, antioxidant status, and gut health of broiler chickens challenged with Eimeria species.
Yadav Sudhir,Teng Po-Yun,Souza Dos Santos Tatiane,Gould Rebecca Lizabeth,Craig Steven Wesley,Lorraine Fuller Alberta,Pazdro Robert,Kim Woo Kyun
Supplementation of broiler diets with feed additives such as chemotherapeutic drugs and antibiotics has side effects, meat residues, and antibiotics resistance complications. Plant-derived natural compounds could be safe and easy substitutes for chemical additives. One of the natural compounds is curcumin, the extract from herbal plant Curcuma longa, known for its antioxidant and antimicrobial properties which may be effective in reducing coccidia infection in poultry. The objective of this study was to evaluate the effects of curcumin on Eimeria challenged (C) and nonchallenged (NC) Cobb 500 broilers. A total of 360 12-day-old male chicks were housed in 36 cages in a completely randomized design with 6 replicates per treatment of 10 birds each cage. The six corn-soybean meal-based treatment diets were fed from day 12 to 20 to C and NC birds in 3-by-two factorial arrangement: nonchallenged control (NCC), NC + 100 mg/kg curcumin, NC + 200 mg/kg curcumin, challenged control (CC), C + 100 mg/kg curcumin, and C + 200 mg/kg curcumin. Broilers in C groups were inoculated orally with 50,000 oocysts of Eimeria maxima, 50,000 oocysts of Eimeria tenella, and 250,000 oocysts of Eimeria acervulina on day 14. The intestinal permeability (day 19), growth performance parameters, and intestinal lesion scoring were measured and recorded on day 20. The means were subjected to two-way ANOVA, and main factors effect and their interactions were considered. The growth performance and permeability were higher (P < 0.001) in the NC and C groups, respectively. However, no interaction was observed between curcumin dose and cocci challenge on both of these parameters. Results from lesion scores and oocyst shedding showed reduction (P < 0.050) in birds fed C + 200 mg/kg curcumin compared with those fed C + 100 mg/kg curcumin or CC. Curcumin treatment showed higher production of GSH (P = 0.002) and total glutathione (GSH+2GSSG) (P = 0.002) but lower GSH/GSSG ratio (P < 0.001) than the NCC group. Curcumin exhibited some positive responses on antioxidant capacity, lesion score, and oocyst shedding in the present study, suggesting that curcumin alone or a combination with other feed additives could be a dietary strategy to improve gut health in broilers.
Immunomodulatory and antiparasitic effects of garlic extract on Eimeria vermiformis-infected mice.
Khalil Atef Mohammed,Yasuda Masahiro,Farid Ayman Samir,Desouky Mohamed Ibrahim,Mohi-Eldin Mouchira Mohammed,Haridy Mohie,Horii Yoichiro
We investigated the immunomodulatory and parasiticidal effects of garlic extract on coccidiosis caused by Eimeria vermiformis infection in male ICR mice. One group received garlic extract daily until the end of the experiment by the oral route from 10 days prior to oral infection with 300 sporulated E. vermiformis oocysts (infected-garlic(+)). The other group served as a control positive with E. vermiformis infection alone (infected-garlic(-)). In the infected-garlic(+) group, garlic extract treatment induced a significant reduction in fecal oocyst output when compared with the infected-garlic(-) group. Histopathological, immunohistochemical, and gene expression analysis for inflammatory cytokines in ileal tissues showed that the garlic extract treatment impaired intracellular development of E. vermiformis during the early stages by increasing the number of intraepithelial CD8(+) T cells and decreasing IL-10 expression. This induced cell cytotoxicity which was reflected by a decrease in oocyst numbers in the intestinal villi and the feces, indicating anticoccidial effects of the garlic extract. However, further studies to explore the precise mechanism of the observed effects of garlic treatment during Eimeria infection are needed to verify our results.
The Effects of Selenium-Enriched Probiotics on Growth Performance, Oocysts Shedding, Intestinal Cecal Lesion Scores, Antioxidant Capacity, and mRNA Gene Expression in Chickens Infected with Eimeria tenella.
Mengistu Berhe Mekonnen,Bitsue Habtom Kiros,Huang Kehe
Biological trace element research
This study was carried out to investigate the effects of selenium-enriched probiotics (SP) supplementation on growth performance, oocysts shedding, intestinal lesions and antioxidant capacities, and mRNA gene expression of local Chinese yellow male chickens infected with Eimeria tenella. One-day-old 270 chickens were randomly assigned into five groups, each consisting of three replicates with 18 chickens per replicate. Chickens in the negative and positive controls (NC, PC, respectively) received basal diets only (0.11 mg Se/kg), whereas the other groups were supplied basal diets with probiotics and designated as (P, 0.11 mg Se/kg), sodium selenite (SS, 0.41 mg Se/kg), and (SP, 0.41 mg Se/kg) groups. At 21 days of age, except the NC group, all other groups were infected by oral gavage with 1.5 × 10 sporulated E. tenella oocysts per chicken. Three chickens were randomly selected from each group for serum, liver, and cecal specimen collection. The results showed that P, SS, and SP had significant increase weight gain and feed intake. Additionally, these groups showed higher activities of serum superoxide dismutase (SOD) and glutathione peroxidase-1 (GPx1) compared to the PC group, whereas feed conversion ratio (FCR), serum catalase (CAT) activity, and malondialdehyde (MDA) content remained lower. Moreover, P, SS, and SP groups had lower oocyst shedding and cecal lesion scores. Significant upregulation of the glutathione peroxidase-1 (GPx1), glutathione peroxidase-4 (GPx4), Selenium W (SelW), and interferon gamma (IFN-γ) mRNA expression were detected in the SP group, which was then followed by SS when compared to the P group, whereas mRNA expression down-regulated in the PC group compared to NC, P, SS, and SP. In the NC and P groups, there were no significant differences in mRNA expression, except that IFN-γ mRNA level upregulated in P. We concluded that selenium-enriched probiotic supplementation has profound effects in enhancing the growth performance, antioxidant capacities, mRNA gene expression, reduced of oocysts shedding, and the cecal lesion scores of chickens and do provide protection against E. tenella.
Recombinant invasive Lactobacillus plantarum expressing the Eimeria tenella fusion gene TA4 and AMA1 induces protection against coccidiosis in chickens.
Liu Yang,Jiang Yan-Long,Liu Jing,Gao Xing,Zhang Zan,Huang Hai-Bin,Yang Wen-Tao,Shi Chun-Wei,Cao Xin,Yang Gui-Lian,Wang Chun-Feng
Coccidiosis is an intestinal parasitic disease that is caused by Eimeria tenella and other species, and it seriously restricts the economic development of the broiler breeding industry. In this study, a recombinant Lactobacillus plantarum with an invasive effect was constructed, and it expressed the TA4-AMA1 protein of E. tenella. After oral immunization with recombinant L. plantarum, specific humoral and mucosal immune levels were measured by indirect ELISA, and the differentiation of T cells was analysed by flow cytometry. After challenge with sporulated oocysts, the body weight, oocyst shedding and cecum lesions of the chicken were evaluated. The results indicated that chickens immunized with recombinant invasive L. plantarum produced higher levels of specific antibodies in the serum than did the non-immunized controls, and the secretory IgA (sIgA) levels were increased in the intestinal washes compared to those of the controls (P < 0.05). Flow cytometry showed that recombinant invasive L. plantarum significantly stimulated T cell differentiation compared to the PBS group (P < 0.01, P < 0.001), and a higher proportion of CD4 and CD8 T cells were detected in peripheral blood. Moreover, the lesion scores and histopathological caecum sections showed that immunizing chickens with recombinant invasive L. plantarum can significantly relieve pathological damage in the cecum (P < 0.01), and the relative body weight gain was 89.64 %, which was higher than the 79.83 % gain in the chickens immunized with non-invasive L. plantarum. After the challenge, faeces from ten chickens in each group were collected between 4 and 7 days, and the oocysts per gram (OPG) was determined by the McMaster technique. The data indicated that oocysts in the faeces of chickens immunized with the recombinant invasive L. plantarum were significantly lower than those of the controls (P < 0.01). The results suggest that recombinant invasive L. plantarum effectively activated immune responses against E. tenella infection and can be used as a candidate vaccine against E. tenella infection.
Immune response and protective efficacy of recombinant Enterococcus faecalis displaying dendritic cell-targeting peptide fused with Eimeria tenella 3-1E protein.
Chen Wenjing,Ma Chunli,Wang Dian,Li Guanghao,Ma Dexing
Avian coccidiosis causes significant economic losses on the global poultry breeding industry. Exploration of new-concept vaccines against coccidiosis has gradually become a research hotspot. In this study, an Enterococcus faecalis strain (MDXEF-1) showing excellent performance isolated from chicken intestinal tract was used as a vector to deliver Eimeria target protein. The plasmid pTX8048-SP-DCpep-NAΔ3-1E-CWA harboring dendritic cell-targeting peptide (DCpep) fusion with Eimeria tenella NAΔ3-1E gene (3-1E protein-coding gene without start codon ATG and terminator codon TAA) was electrotransformed into MDXEF-1 to generate the recombinant bacteria MDXEF-1/pTX8048-SP-DCpep-NAΔ3-1E-CWA in which NAΔ3-1E protein was covalently anchored to the surface of bacteria cells by cell wall anchor (CWA) sequence. The expression of target fusion protein DCpep-NAΔ3-1E-CWA was detected by Western blot. Each chicken was immunized 3 times at 2-wk intervals with live E. faecalis expressing DCpep-NAΔ3-1E fusion protein (DCpep-NAΔ3-1E group), live E. faecalis expressing NAΔ3-1E protein (NAΔ3-1E group), and live E. faecalis containing empty vector only. The 3 immunized groups were then challenged with homologous E. tenella sporulated oocyst after immunizations, and the immune response and protective efficacy in each group were evaluated. The results showed that serum IgG levels, secretory IgA levels in cecal lavage, proportion of CD4+ and CD8α+ cells in peripheral blood, and mRNA expression levels of IL-2 and IFN-γ in the spleen were significantly higher in chickens in the DCpep-NAΔ3-1E group than in chickens of the NAΔ3-1E group (P < 0.05). Oral immunization to chickens with live E. faecalis expressing DCpep-NAΔ3-1E offered more protective efficacy against homologous challenge including significant improved body weight gain, increased oocyst decrease ratio, and reduced average lesion scores in cecum compared with chickens with live E. faecalis expressing NAΔ3-1E protein. These results suggest that recombinant E. faecalis expressing dendritic cell-targeting peptide fusion with E. tenella 3-1E protein could be a potential approach for prevention of Eimeria infection.
The effect of a Local isolate and Houghton strain of on clinical and growth parameters following challenge in chickens vaccinated with IMMUCOX and LIVACOX vaccines.
Akanbi Olatunde B,Taiwo Victor O
Journal of parasitic diseases : official organ of the Indian Society for Parasitology
Coccidiosis, a parasitic infection caused by one or more of the numerous species of is recognised as the disease that has severe economic impact on poultry production. In an experiment to determine the effect of vaccination on the infection with a Local isolate and Houghton strain of (). Ninety (90) day-old Dominant black cockerel chicks of D109 strain were randomly divided into nine groups (A-I). Two commercially available anti-coccidial vaccines-Livacox (Biopharm, Czech Republic) and Immucox (Vetech, Ontario, Canada) were used to immunize the chicks. Immunization was done at 5 days-old by oral gavage and infection was carried out with 1.7 × 10 of either Houghton strain or a field (Local) isolate at 4-weeks old. Six (6) days post-challenge, generalized pallor, moderate bloody diarrhea, slight reduction in feed intake and weight gain characterized the infected groups. There is a significant difference ( < 0.05) in packed cell volume of group B, unvaccinated and infected with Houghton strain of and group E vaccinated with Livacox and infected with Houghton strain. The Immunogenicity of Livacox and Immucox was determined primarily on the ability of the vaccinated chickens to overcome the effect of the virulent challenge by including blood loss, reduction in feed intake and feed conversion and weight loss. The two anticoccidial vaccines used in this experiment were effective in varying degrees especially with the local isolate.
Eimeria tenella Eimeria-specific protein that interacts with apical membrane antigen 1 (EtAMA1) is involved in host cell invasion.
Li Cong,Zhao Qiping,Zhu Shunhai,Wang Qingjie,Wang Haixia,Yu Shuilan,Yu Yu,Liang Shashan,Zhao Huanzhi,Huang Bing,Dong Hui,Han Hongyu
Parasites & vectors
BACKGROUND:Avian coccidiosis is a widespread, economically significant disease of poultry, caused by several Eimeria species. These parasites have complex and diverse life-cycles that require invasion of their host cells. This is mediated by various proteins secreted from apical secretory organelles. Apical membrane antigen 1 (AMA1), which is released from micronemes and is conserved across all apicomplexans, plays a central role in the host cell invasion. In a previous study, some putative EtAMA1-interacting proteins of E. tenella were screened. In this study, we characterized one putative EtAMA1-interacting protein, E. tenella Eimeria -specific protein (EtEsp). METHODS:Bimolecular fluorescence complementation (BiFC) and glutathione S-transferase (GST) fusion protein pull-down (GST pull-down) were used to confirm the interaction between EtAMA1 and EtEsp in vivo and in vitro. The expression of EtEsp was analyzed in different developmental stages of E. tenella with quantitative PCR and western blotting. The secretion of EtEsp protein was tested with staurosporine when sporozoites were incubated in complete medium at 41 °C. The localization of EtEsp was analyzed with an immunofluorescence assay (IFA). An in vitro invasion inhibition assay was conducted to assess the ability of antibodies against EtEsp to inhibit cell invasion by E. tenella sporozoites. RESULTS:The interaction between EtAMA1 and EtEsp was confirmed with BiFC and by GST pull-down. Our results show that EtEsp is differentially expressed during distinct phases of the parasite life-cycle. IFA showed that the EtEsp protein is mainly distributed on the parasite surface, and that the expression of this protein increases during the development of the parasite in the host cells. Using staurosporine, we showed that EtEsp is a secreted protein, but not from micronemes. In inhibition tests, a polyclonal anti-rEtEsp antibody attenuated the capacity of E. tenella to invade host cells. CONCLUSION:In this study, we show that EtEsp interacts with EtAMA1 and that the protein is secreted protein, but not from micronemes. The protein participates in sporozoite invasion of host cells and is maybe involved in the growth of the parasite. These data have implications for the use of EtAMA1 or EtAMA1-interacting proteins as targets in intervention strategies against avian coccidiosis.
Research Note: Correlation analysis of interleukin-6, interleukin-8, and C-C motif chemokine ligand 2 gene expression in chicken spleen and cecal tissues after Eimeria tenella infection in vivo.
Yu Hailiang,Zou Wenbin,Wang Xiaohui,Dai Guojun,Zhang Tao,Zhang Genxi,Xie Kaizhou,Wang Jinyu,Shi Huiqiang
IL-6, IL-8, and C-C motif chemokine ligand 2 (CCLi2) are important factors in inflammatory and immune responses. To investigate their relationships in the spleen and cecum and between coccidiosis-infected and uninfected states, we performed quantitative real-time PCR to compare the relative expression difference of IL-6, IL-8, and CCLi2 in the same tissues between the infection and control groups. In addition, the correlations of the relative expression levels of these 3 genes were determined in the same and different tissues within the same group. The results showed that the expression levels of IL-6, IL-8, and CCLi2 in the spleen and cecum of the infected group were all higher than those of the uninfected group (P < 0.05). The correlation coefficients among the IL-6, IL-8, and CCLi2 expression levels in the spleen or cecum were all positive in both the infection and control groups. In the spleen tissues, CCLi2 expression was strongly correlated with IL-6 and IL-8 in the uninfected group (P < 0.01), and the correlation coefficients reached 0.853 (R = 0.728) and 0.996 (R = 0.992), respectively. The expression of CCLi2 was also strongly correlated with IL-8 (R reached 0.890, R = 0.792) in the infected group. In the cecal tissues, the expression levels of the 3 genes were all extremely significantly correlated in the uninfected group (P < 0.01), and the correlation coefficients ranged from 0.498 to 0.765, indicating moderate correlations. The expression of IL-6 was extremely significantly positively correlated with IL-8 and CCLi2 in the infected group (P < 0.01), with moderate correlations (R ranged from 0.469-0.639). In addition, the expression levels of the 3 genes were not significantly correlated (P > 0.05) between the spleen and cecum tissues in either the infection group or the control group. These results indicate that IL-6, IL-8, and CCLi2 were correlated and play an important role in coccidiosis infection of Jinghai yellow chicken. Our data also provide a basis for further exploring the role of these 3 genes in genetic breeding for coccidiosis resistance.
Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system.
Tang Xinming,Suo Jingxia,Liang Lin,Duan Chunhui,Hu Dandan,Gu Xiaolong,Yu Yonglan,Liu Xianyong,Cui Shangjin,Suo Xun
Eimeria tenella has emerged as valuable model organism for studying the biology and immunology of protozoan parasites with the establishment of the reverse genetic manipulation platform. In this report, we described the application of CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (endonuclease) system for efficient genetic editing in E. tenella, and showed that the CRISPR/Cas9 system mediates site-specific double-strand DNA breaks with a single guide RNA. Using this system, we successfully tagged the endogenous microneme protein 2 (EtMic2) by inserting the red fluorescent protein into the C-terminal of EtMic2. Our results extended the utility of the CRISPR/Cas9-mediated genetic modification system to E. tenella, and opened a new avenue for targeted investigation of gene functions in apicomplexan parasites.
iTRAQ-based comparative proteomic analysis of cells infected with Eimeria tenella sporozoites.
Zhao Zongping,Zhao Qiping,Zhu Shunhai,Huang Bing,Lv Ling,Chen Ting,Yan Ming,Han Hongyu,Dong Hui
Parasite (Paris, France)
Eimeria tenella is an obligate intracellular parasite that actively invades cecal epithelial cells of chickens. When E. tenella infects a host cell, the host produces a corresponding change to deal with damage caused by this infection. To date, our knowledge on the mechanism of how the host cell responds to E. tenella infection is highly limited at both the molecular and cellular levels. In this study, isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS was used to screen the differentially expressed proteins (DEPs) in BHK-21 cells infected with E. tenella sporozoites for 24 h post infection. In total, 6139 non-redundant distinct proteins were identified and 195 of these were found to have a fold change ratio ≥1.3 or ≤0.7 and p < 0.05, including 151 up-regulated proteins and 44 down-regulated proteins. The reliability of the proteomic data was further validated with qPCR and western blot. Gene Ontology enrichment indicated that the up-regulated DEPs were mainly involved in binding and catalytic activity, whereas the down-regulated DEPs were catalytic activity and molecular function regulators. Furthermore, KEGG pathway analysis showed that the DEPs participated in the PI3K-Akt, chemokine, Ras, Wnt, and p53 signaling pathways and so on, and the up-regulated and down-regulated DEPs mainly related to the ribosome and mRNA surveillance pathway, respectively. The data in this study provide an important basis to further analyze E. tenella host cell interactions.
Protective effects of a food-grade recombinant Lactobacillus plantarum with surface displayed AMA1 and EtMIC2 proteins of Eimeria tenella in broiler chickens.
Liu Qiong,Jiang Yanlong,Yang Wentao,Liu Yongshi,Shi Chunwei,Liu Jing,Gao Xing,Huang Haibin,Niu Tianming,Yang Guilian,Wang Chunfeng
Microbial cell factories
BACKGROUND:Avian coccidiosis posts a severe threat to poultry production. In addition to commercial attenuated vaccines, other strategies to combat coccidiosis are urgently needed. Lactobacillus plantarum has been frequently used for expression of foreign proteins as an oral vaccine delivery system using traditional erythromycin resistance gene (erm). However, antibiotic selection markers were often used during protein expression and they pose a risk of transferring antibiotic resistance genes to the environment, and significantly restricting the application in field production. Therefore, a food-grade recombinant L. plantarum vaccine candidate would dramatically improve its application potential in the poultry industry. RESULTS:In this study, we firstly replaced the erythromycin resistance gene (erm) of the pLp_1261Inv-derived expression vector with a non-antibiotic, asd-alr fusion gene, yielding a series of non-antibiotic and reliable, food grade expression vectors. In addition, we designed a dual-expression vector that displayed two foreign proteins on the surface of L. plantarum using the anchoring sequences from either a truncated poly-γ-glutamic acid synthetase A (pgsA') from Bacillus subtilis or the L. acidophilus surface layer protein (SlpA). EGFP and mCherry were used as marker proteins to evaluate the surface displayed properties of recombinant L. plantarum strains and were inspected by western blot, flow cytometry and fluorescence microscopy. To further determine its application as oral vaccine candidate, the AMA1 and EtMIC2 genes of E. tenella were anchored on the surface of L. plantarum strain. After oral immunization in chickens, the recombinant L. plantarum strain was able to induce antigen specific humoral, mucosal, and T cell-mediated immune responses, providing efficient protection against coccidiosis challenge. CONCLUSIONS:The novel constructed food grade recombinant L. plantarum strain with double surface displayed antigens provides a potential efficient oral vaccine candidate for coccidiosis.
Prolonging and enhancing the protective efficacy of the EtMIC3-C-MAR against eimeria tenella through delivered by attenuated salmonella typhimurium.
Zhao Ningning,Lv Junfeng,Lu Yaru,Jiang Yingying,Li Hongmei,Liu Yumin,Zhang Xiao,Zhao Xiaomin
The microneme adhesive repeats (MAR) of Eimeria tenella microneme protein 3 (EtMIC3) are associated with binding to and invasion of host cells. Adhesion and invasion-related proteins or domains are often strongly immunogenic, immune responses mounted against these factors that play a key role in blocking invasion. In the present study, an oral live vaccine consisting of attenuated Salmonella typhimurium X4550 carrying two MAR domains fragment (St-X4550-MAR) was constructed and its protective efficacies were evaluated. The results showed that St-X4550-MAR was more immunogenic and conferred a higher degree of protection than recombinant MAR polypeptide as reflected by increased body weight, decreased oocyst shedding and lesion scores, increased serum IgG and cecal sIgA antibody production, and increasing levels of interferon-γ and interleukin-10. Thus, MAR domains are highly immunogenic and St-X4550-MAR had moderate activity against E. tenella infection by stimulating humoral, mucosal and cellular immunity. Chickens immunized with our constructed live vaccine provided considerable protections as early as at 10 d post-immunization (ACI: 155.17), and maintained higher protection levels at 20 d post-immunization (ACI: 173.66), and at 30 d post-immunization (ACI: 162.4). While the protective efficacy of chickens immunized with the recombinant MAR peptides showed a decreased trend as the post immunization time prolonging. Thus, using live-attenuated S. typhimurium X4550 as a vaccine expression and delivery system can significantly improve the protective efficacy and duration of protective immunity of MAR of EtMIC3.
The effect of autophagy on the survival and invasive activity of Eimeria tenella sporozoites.
Qi Nanshan,Liao Shenquan,Abuzeid Asmaa M I,Li Juan,Wu Caiyan,Lv Minna,Lin Xuhui,Hu Junjing,Yu Linzeng,Xiao Wenwan,Sun Mingfei,Li Guoqing
Autophagy is a cellular process that is vital for the maintenance of homeostasis in eukaryotic cells. Currently, autophagy-related genes (atgs) in the Eimeria tenella genome database have been reported, but very little is known about the effects of autophagy on the survival and invasive activity of this protozoan. In this study, we investigated the autophagy in E. tenella sporozoites under starvation and autophagy-modulators treatments and evaluated the autophagy influence on cellular adenosine triphosphate (ATP) levels, the survival rate and the invasive activity of the sporozoites. The results showed that the autophagy could be induced in the sporozoites by starvation or inducer rapamycin (RP), but it could be inhibited by 3-methyladenine (3-MA) treatment. The sporozoites after starvation and RP-treatment displayed punctate signals of EtATG8 and formed autophagosomes. The survival rate of the sporozoites under starvation was significantly lower than that in the control group, whereas the ATP levels in sporozoite were far greater than those in the control. The quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) showed that the invasive activity of the sporozoites was up- and down-regulated by RP and 3-MA induction, respectively. Our results indicate that autophagy has effects on the survival and invasive activity of E. tenella sporozoites, which may provide new insights into anti-coccidial drugs.
Optimization of Immunization Procedure for Eimeria tenella DNA Vaccine pVAX1-pEtK2-IL-2 and Its Stability.
Zhang Bucai,Yuan Cheng,Song Xiaokai,Xu Lixin,Yan Ruofeng,Shah Muhammad Ali A,Guo Changming,Zhu Shanyuan,Li Xiangrui
PURPOSE:To seek for the optimal immunization procedure of DNA vaccine pVAX1-pEtK2-IL-2 which was produced via cloning pEtK2 antigen gene of Eimeria tenella (E. tenella) and chicken IL-2 (chIL-2) gene into expression vector pVAX1. METHODS:The doses, routes, times of inoculation and ages of the first inoculation of chickens were optimized. The stability of the vaccine, including store temperature and time, was also explored. The effects of the protective immunity against challenge infection were assessed according to average body weight gain, survival rate, oocyst output, lesion score and the anti-coccidial index (ACI). RESULTS:The results suggested that intramuscular inoculation was the most efficient route to elicit immune response and 80 μg was the optimal immune dose. Two time injections induced more effective protection compared to single injection, the effect of the first injection at 14 days old was optimal. The immune efficacy of the vaccine stored at different time and temperature was very stable. CONCLUSIONS:The optimal immunization procedure for Eimeria tenella DNA vaccine pVAX1-pEtK2-IL-2 is 80 μg DNA, two time injections at 14 and 21 days old, respectively, by intramuscular inoculation.
Elongation Factor-1α (EF-1α) Coadministered with Chicken IL-7 (chIL-7) DNA Vaccine Emulsified in Montanide Gel 01 Adjuvant Enhanced the Immune Response to Infection in Broiler Chickens.
Panebra Alfredo,Lillehoj Hyun S
The current study was undertaken to assess the vaccine efficacy of EF-1α/chicken IL-7 (chIL-7) DNA vaccine when administered with Montanide Gel 01 adjuvant against live challenge in commercial broiler chickens. The criteria used for the evaluation of vaccine efficacy were weight gain, duodenal lesion scores, oocyst counts, humoral antibody response, and duodenal proinflammatory cytokine gene expression. Chickens vaccinated with EF-1α (100 µg)/chIL-7 (20 µg) in Gel 01 PR adjuvant showed body weight gain similar to the uninfected control and higher oocyst shedding, a lower gut lesion score, and higher proinflammatory cytokine gene expression than did the infected controls. Moreover, chickens vaccinated with chIL-7 (20 µg) in Gel 01 PR adjuvant shed fewer oocysts with reduced gut lesion scores and produced higher levels of anti-EF-1α serum antibody than did the infected control. Chickens vaccinated with EF-1α (50 µg)/chIL-7 (20 µg) in Gel 01 PR adjuvant showed higher weight gains than did the infected control and shed significantly fewer oocysts than the infected control. Furthermore, chickens vaccinated with EF-1α (100 µg) in Gel 01 PR adjuvant demonstrated the lowest anti-EF-1α serum antibody levels. This study demonstrated the beneficial effects of using EF-1α and/or host cytokine chIL-7 DNA vaccine together with Gel 01 PR adjuvant to improve T-cell-mediated effector function in broiler chickens challenged with live .
Transitions in morphological forms and rapid development of the asexual schizonts of Eimeria tenella through serial passaging in chicks.
Matsubayashi Makoto,Yamaguchi Hiroki,Hatta Takeshi,Kawahara Fumiya,Hatabu Toshimitsu,Iseki Hiroshi,Yamagishi Junya,Isobe Takashi,Teramoto Isao,Kaneko Akira,Kita Kiyoshi,Tsuji Naotoshi,Sasai Kazumi
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases
Attenuated strains of avian Eimeria parasites, generated by the selection of precocious lines through serial passaging in chicks, have been used widely as live vaccines. Detailed morphological transitions including their life cycle depending on the passages remain poorly understood. Here, we showed early development and acceleration of transitions in morphological forms of the asexual schizonts of E. tenella that had been attenuated for virulence by serial passaging. Our results may be helpful in understanding parasitism, facilitating further molecular analyses such as comparative genomic or transcriptomic tests.
Statistical analyses of chicken intestinal lesion scores in battery cage studies of anti-coccidial drugs.
Kang Qing,Vahl Christopher I,Fan Huihao,Geurden Thomas,Ameiss Keith A,Taylor Lucas P
Establishing the efficacy of an anti-coccidial drug in poultry begins with conducting multiple battery cage studies, where the target animals are challenged with single and mixed Eimeria species inoculum under controlled laboratory conditions. One of the primary outcomes in a battery cage study is the intestinal lesion score defined on a discrete ordinal scale of 0 to 4. So far, the statistical analysis of lesion scores has routinely employed the linear mixed model (LMM). This present work proposes to apply the generalized linear mixed model (GLMM) with the cumulative logit link to statistically analyze coccidial lesion scores collected from battery cage studies. Upon applying this new approach on 9 datasets generated by challenging battery-cage-housed broilers with various mixtures of Eimeria species, it is observed that the GLMM fitted adequately to the data, produced variance component estimates that agreed with the experimental setup, and, at the 0.05 significance level, generated statistical results in complete concordance with the LMM approach. Advantages of the proposed GLMM over the LMM are discussed from several standpoints. Parallel to the regulatory requirement of a ≥1-unit reduction in the mean lesion score for clinical relevant efficacy under the LMM, the clinical relevancy criterion under the GLMM could be set as a ≥10-fold increase in the odds of having low lesion scores. That is, the effect of an anti-coccidial drug product would be deemed clinically relevant in battery-cage studies when the odds of having low lesion scores with the medication is 10 times or more than the odds without the medication.
Effect of 25-hydroxycholecalciferol supplementation on turkey performance and immune cell parameters in a coccidial infection model.
Shanmugasundaram Revathi,Morris Antrison,Selvaraj Ramesh K
This study was conducted to identify the effects of 25-OH cholecalciferol supplementation to turkeys on the immune cells parameters, fecal coccidial oocyst shedding, macrophage nitric oxide production, T regulatory cell cytokine production, and production parameters during a coccidial challenge. A total of 200 1-day-old turkey poults were supplemented with 27.5, 55, 82.5, or 110 μg/kg 25-OH cholecalciferol and challenged or not challenged with coccidial oocysts in a 4 × 2 factorial set up of treatments. Birds fed 110 μg/kg of 25-OH cholecalciferol and infected with coccidial oocyst had 41% lower (P < 0.05) fecal oocyst and 53% higher (P < 0.05) macrophage nitric oxide production than the birds fed 27.5 μg/kg of 25-OH cholecalciferol and infected with coccidial oocyst at 5 d post-coccidial infection. Birds fed 82.5 μg/kg 25-OH cholecalciferol and infected with coccidial oocyst had 5-fold higher (P < 0.05) IL-1 mRNA amounts than the birds fed 27.5 μg/kg of 25-OH cholecalciferol and infected with coccidial oocyst. Birds fed 110 μg/kg 25-OH cholecalciferol and infected with coccidial oocyst had 5.3-fold higher (P < 0.05) IL-10 mRNA amounts than the birds fed 27.5 μg/kg of 25-OH cholecalciferol and infected with coccidial oocyst at 5 d post-coccidial infection. CD4+CD25+ cells from birds fed 110 μg of 25-OH cholecalciferol and infected with coccidial oocyst had 12-fold higher (P < 0.05) IL-10 mRNA than that from the birds fed 27.5 μg/kg of 25-OH cholecalciferol and infected with coccidial oocyst. In conclusion, supplementing birds with 101 μg/kg 25-OH cholicalciferol decreases coccidial oocyst shedding in the feces and could be a nutritional strategy to reduce the coccidial infection and spread in turkeys.
Efficacy of dietary supplemental insoluble fibrous materials in ameliorating adverse effects of coccidial challenge in broiler chickens.
Sadeghi Amin,Toghyani Majid,Tabeidian Sayed Ali,Foroozandeh Amir Davar,Ghalamkari Gholamreza
Archives of animal nutrition
The current experiment was designed to examine effects of dietary supplemental sunflower hulls (SH) and rice hulls (RH) on growth performance, carcass traits, intestinal morphology, lesion score and oocyst shedding in broiler chickens exposed to coccidial challenge. A total of 540 broiler chickens (Ross 308) were assigned to six dietary treatments based on a factorial arrangement (2 × 3) across 1-14, 14-28 and 28-42-d periods. Experimental treatments consisted of broiler chickens without or with coccidial challenge each offered with three different diets: a basal diet or basal diet supplemented with either RH or SH at 40 g/kg diet, respectively. Infection with impaired daily weight gain (DWG) and feed conversion ratio (FCR) of broiler chickens during growing period ( < 0.05) while supplementation of SH or RH reduced the adverse effect of coccidiosis so that birds had similar DWG to those fed the basal diet without infection. However, only dietary SH improved the FCR of broilers challenged with coccidiosis. Regardless of coccidial challenge, dietary access to insoluble fibre improved performance of broilers across the growing period ( < 0.05); however, this effect was not observed during the entire rearing period. Relative weights of liver and pancreas were increased in birds subjected to coccidial challenge on d 21 of age ( < 0.05). Moreover, relative weights of the intestinal segments were enhanced ( < 0.05). Furthermore, gizzard weights were higher in birds receiving diets added with fibre ( < 0.05). Infection with coccidiosis decreased villus height and villus height to crypt depth ratio in duodenum of broilers which received the basal diet compared with those fed the same feed without coccidial challenge ( < 0.05). However, supplemental SH could decrease the negative effect of infection on the noted intestinal morphometric attributes. Similarly, a marked reduction was observed for lesion score and faecal oocyst excretion of challenged broilers fed on dietary supplemental fibre ( < 0.05). In conclusion, supplementation of insoluble fibre could ameliorate negative effects of coccidial challenge on DWG of broiler chickens and inclusion of SH in diet of birds exposed to infection could be recommended.
Stable transfection of Eimeria necatrix through nucleofection of second generation merozoites.
Duan Chunhui,Hu Dandan,Tang Xinming,Suo Jingxia,Wang Si,Zhang Sixin,Tao Geru,Li Chao,Wang Chaoyue,Gu Xiaolong,Tang Xiaoli,Huang Guangping,Xiang Biqi,Wu Shaoqiang,Mamoun Choukri Ben,Suo Xun,Liu Xianyong
Molecular and biochemical parasitology
Eimeria spp., the causative agents of coccidiosis, are the most common protozoan pathogens of chickens. Infection with these parasites can result in poor development or death of animals leading to a devastating economic impact on poultry production. The establishment of transfection protocols for genetic manipulation of Eimeria species and stable expression of genes would help advance the biology of these parasites as well as establish these organisms as novel vaccine delivery vehicles. Here, we report the selection of the first stable transgenic E. necatrix population, EnHA1, consitutively expressing the EYFP reporter following transfection of the 2 generation merozoites with a linear DNA fragment harboring the EYFP reporter gene, the HA1 gene from the avian influenza virus H9N2 and the TgDHFR-TS selectable marker, which confers resistance to pyrimethamine. Transfected merozoites were inoculated into chickens via the cloacal route, and feces from 18 h to 72 h post inoculation were collected and subjected to subsequent serial passages, FACS sorting and pyrimethamine selection. A gradual increase in the number of EYFP-expressing sporulated oocysts was noticed with more than 90% EYFP + oocysts obtained after five passages. Immunofluorescence assay confirmed successful expression of the HA1 antigen in the EnHA1 population. The ability to genetically manipulate E. necatrix merozoites and express heterologous genes in this parasite will pave the way for possible use of this organism as a vaccine-delivery vehicle.
Evaluation of herbal coccidiostat 'Coxynil' in broiler.
Kurkure N V,Kolte S W,Bhandarkar A G,Kalorey D R
Indian journal of experimental biology
Anticoccidial efficacy of "Coxynil" a polyherbal preparation was tested against Eimeria tenella in broilers. Body weight of birds challenged with E. tenella in Coxynil treated groups was higher as compared to Coxynil untreated. Oocyst out put, lesion score, HI titres against New Castle disease virus were significantly higher in Coxynil supplemented groups in comparison to Coxynil un-supplemented groups. Examination of ceaca of the birds, revealed that the Coxynil interfered with life cycle of coccidia. The typical second generation schizonts were absent in ceacal section of Coxynil treated groups. The results indicate that Coxynil is effective herbal coccidiostat.
Anticoccidial Effect of Herbal Powder "Shi Ying Zi" in Chickens Infected with .
Song Xu,Li Yunhe,Chen Shufan,Jia Renyong,Huang Yongyuan,Zou Yuanfeng,Li Lixia,Zhao Xinxin,Yin Zhongqiong
Animals : an open access journal from MDPI
Coccidiosis is one of the most economically important diseases affecting the poultry industry. Currently, anticoccidial drugs used in veterinary clinics show many deficiencies, and new control measures are urgently needed. This study presents an anticoccidial herbal powder "Shi Yin Zi", which consists of (L.) Cuss, Hand.-Mazz., and sodium chloride. In chickens infected with , supplementation with "Shi Yin Zi" powder for 3 d prior to infection or treatment with "Shi Yin Zi" powder after infection could improve the survival rate and relative growth rate and alleviate the pathological changes in the cecum, liver, and kidney. "Shi Yin Zi" powder could recover the levels of alanine aminotransferase, creatinine, albumin, and triglycerides in serum. The hemorrhage occurrence and total number of oocysts in feces were reduced. The anti-coccidial indexes reached 165 for the prophylactic effect and 144 for the therapeutic effect. The anti-coccidial effects were equal to positive controls (monensin and sulfamlopyrazine). These results suggest that "Shi Ying Zi" powder possesses a potent anticoccidial effect and exhibits the potential to control infection.
Herbal Remedies for Coccidiosis Control: A Review of Plants, Compounds, and Anticoccidial Actions.
Muthamilselvan Thangarasu,Kuo Tien-Fen,Wu Yueh-Chen,Yang Wen-Chin
Evidence-based complementary and alternative medicine : eCAM
Coccidiosis is the bane of the poultry industry causing considerable economic loss. Eimeria species are known as protozoan parasites to cause morbidity and death in poultry. In addition to anticoccidial chemicals and vaccines, natural products are emerging as an alternative and complementary way to control avian coccidiosis. In this review, we update recent advances in the use of anticoccidial phytoextracts and phytocompounds, which cover 32 plants and 40 phytocompounds, following a database search in PubMed, Web of Science, and Google Scholar. Four plant products commercially available for coccidiosis are included and discussed. We also highlight the chemical and biological properties of the plants and compounds as related to coccidiosis control. Emphasis is placed on the modes of action of the anticoccidial plants and compounds such as interference with the life cycle of Eimeria, regulation of host immunity to Eimeria, growth regulation of gut bacteria, and/or multiple mechanisms. Biological actions, mechanisms, and prophylactic/therapeutic potential of the compounds and extracts of plant origin in coccidiosis are summarized and discussed.
Comparison of the anticoccidial effect of granulated extract of Artemisia sieberi with monensin in experimental coccidiosis in broiler chickens.
Pirali Kheirabadi Khodadad,Kaboutari Katadj Jahangir,Bahadoran Shahab,Teixeira da Silva Jaime A,Dehghani Samani Amir,Cheraghchi Bashi Mehdi
Coccidiosis is the most important parasitic disease of the poultry production industry. Due to increasing resistance to conventional anticoccidial agents, it is necessary to find new anticoccidial compounds. Herbal compounds such as those from Artemisia species are promising weapons in this regard since preliminary studies have shown its anticoccidial effects. To compare the anticoccidial effect of a granulated extract of Artemisia sieberi (GEAS) versus monensin in experimental broiler coccidiosis, 120 one-day old Ross 308 broiler chickens were divided in four groups, each with three replicates (n=10). Group 1 was separated as an uninfected negative control and received no treatment. At 21days of age, groups 2, 3 and 4 were inoculated with a mixed suspension of 2×10(5) oocysts of Eimeria tenella, Eimeria maxima, Eimeria acervulina and Eimeria necatrix. Group 2 was maintained as an infected positive control and received no treatment while groups 3 and 4 received GEAS (5mg/kg feed), and monensin (110mg/kg feed) from the first day until 42days of age as a feed additive, respectively. Five days after inoculation, the number of oocycts per gram (OPG) of feces for 7 successive days was measured. Also, mean body weight (MBW), weight gain (WG), feed intake (FI) and feed conversion ratio (FCR) were determined weekly in all groups and replicates. The results showed that GEAS and monensin improved performance attributes (FI, MBW, WG, FCR) and significantly (P<0.05) decreased OPG in inoculated broiler chickens.
Prophylactic and therapeutic effects of a novel granulated formulation of Artemisia extract on broiler coccidiosis.
Kaboutari Jahangir,Arab Hossien Ali,Ebrahimi Kambiz,Rahbari Sadegh
Tropical animal health and production
PURPOSE:Coccidiosis is an important parasitic disease in poultry industry. Owing to the development of drug resistance against Eimeria and concerns about drug residues, attentions toward the alternative compounds including herbal medicines have been increased. This study aimed to examine the prophylactic and therapeutic effects of a new formulation derived from Artemisia sieberi extract on avian coccidiosis. METHODS:The extract was obtained from A. sieberi using petroleum ether, and then, it was formulated into a wet granule. Three hundred and sixty 1-day-old broilers were divided into six groups, each with three replicates (n = 20); the first group was chosen as the noninfected control group and the remainders were challenged by oral administration of 250,000 oocysts of Eimeria tenella per chick on day 21. Group 2 (as nontreated control) received no treatment, but group 3 (as prophylactic group) received 0.5 mg/kg artemisinin 10 days before challenging with E. tenella. However, groups 4, 5, and 6 received 1 mg/kg artemisinin on the next day and at 24 and 48 h after the challenge, respectively, for 5 days. Mortality rate, bloody diarrhea score, lesion score, and oocysts per gram (OPG) of feces were used to determine the anticoccidial effects of the formulation. RESULTS:The granule significantly (p < 0.05) reduced mortality, diarrhea, and lesion score in the treated groups. The OPG output was also significantly (p < 0.0001) reduced in the groups that received formulation. However, there was no significant difference between the prophylactic and therapeutic groups. CONCLUSION:The results suggested that this new formulation is a promising herbal medicine that can be used as a prophylactic or therapeutic product to control avian coccidiosis.
Efficacy of a commercial herbal formula in chicken experimental coccidiosis.
Pop Loredana Maria,Varga Erzsébet,Coroian Mircea,Nedișan Maria E,Mircean Viorica,Dumitrache Mirabela Oana,Farczádi Lénárd,Fülöp Ibolya,Croitoru Mircea Dumitru,Fazakas Mihaly,Gyӧrke Adriana
Parasites & vectors
BACKGROUND:Coccidiosis represents a serious threat to the poultry industry, affecting production and causing high morbidity, mortality and significant costs resulting from treatment and prophylaxis. In-feed anticoccidials have been used for decades for managing avian coccidiosis and were very effective until drug resistance emerged. The use of natural remedies has become a promising alternative in combating coccidiosis in chickens. Therefore, the purpose of the present study was to assess the efficiency of a commercial herbal formula (H), as oral liquid preparations, in experimental chicken coccidiosis. METHODS:Two independent controlled battery experiments (BE1 and BE2) were designed and the product was tested in 3 different formulas (H1, H2 and H3): H1 contained a propylene glycol extract of Allium sativum and Thymus serpyllum; H2 contained Origanum vulgare, Satureja hortensis and Chelidonium majus; and H3 contained Allium sativum, Urtica dioica, Inula helenium, Glycyrrhiza glabra, Rosmarinus officinalis, Chelidonium majus, Thymus serpyllum, Tanacetum vulgare and Coriandrum sativum. Chickens were divided into five groups for each BE as follows: (i) uninfected untreated control (UU1, UU2); (ii) infected untreated control (IU1, IU2); (iii) infected treated with amprolium (ITA1, ITA2); and (iv, v) two experimental groups infected treated with H1 (ITH1) and H2 (ITH2) formulas in the BE1 and with H3 (ITH3-5 and ITH3-10) formula in the BE2. The chickens from infected groups were challenged with 5000 (BE1) and 50,000 (BE2) sporulated oocysts of Eimeria spp. (E. acervulina, E. tenella and E. maxima), respectively. The anticoccidial efficacy was assessed by recording the following: oocysts output (OPG), lesion score (LS), weight gain (WG), feed conversion ratio (FCR) and anticoccidial index (ACI). Additionally, polyphenolics and flavonoids (caffeic-chlorogenic acid, apigenin, kaempferol, luteolin, quercitin, quercitrin) from herb extracts found in H3 formula were determined by the liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. RESULTS:H1 and H2 reduced the WG, and increased the FCR and OPG compared with controls. H1 reduced the duodenal lesions, whilst H2 reduced the caecal lesions, compared with control. H3 decreased the OPG of Eimeria spp., reduced the total lesion score and improved the zootechnical performance (weight gain and feed conversion ratio). According to ACI value, H1 and H2 had no efficacy on Eimeria spp. infection, but H3 had good to marked anticoccidial effect, the ACI being slightly greater in the group ITH3-5. According to the results of LC-MS/MS, the concentration of polyphenols in H3 formula was the highest, the sum of chlorogenic acid and caffeic acid being 914.9 µg/ml. CONCLUSIONS:H3 formula is a promising natural anticoccidial and field trials are recommended in order to validate the obtained data.
Anticoccidial effects of Artemisia annua ethanolic extract: prevention, simultaneous challenge-medication, and treatment.
Fatemi Ahmadreza,Asasi Keramat,Razavi Seyed Mostafa
The effect of Artemisia annua ethanolic extract (AE) as a potential source of herbal anticoccidial activity was investigated on experimental coccidiosis in chicken. One hundred ninety-two one-day-old chicks were divided in to 8 groups (n = 24) including AE prevention group, AE-treated group, simultaneously challenged AE-medicated group, challenged-untreated group (positive control), unchallenged-untreated group (negative control), salinomycine prevention group, salinomycine-treated group, and simultaneously challenged salinomycine-medicated group, in a completely randomized design. Oral challenge carried out by a suspension containing a mixture of 200,000 oocysts Eimeria acervulina, 30,000 oocysts Eimeria necatrix, and 20,000 oocysts Eimeria tenella on day 21 of age. Weight gain in AE prevention group significantly increased compared to positive control group (p < 0.05). Unlike salinomycine prevention group, the food conversion ratio (FCR) of AE prevention group was not significantly higher than negative control. Oocyst per gram (OPG) in simultaneously challenged AE-medicated group had no significant difference, while for 38% of the days, in simultaneously challenged salinomycine-medicated group significantly decreased (p < 0.05). The food intake of AE-treated group had no significant difference with salinomycine-treated group (p > 0.05). In half of the days of OPGs sampling, AE-treated group was reduced significantly compared to positive control group (p < 0.05). Collectively, the in vivo study of anticoccidial effects of AE in the prevention section was more effective than the treatment section, while the treatment section was more effective than the simultaneous section. We concluded that AE has a potential value to use as an herbal medicine for preventive measure in chicken coccidiosis.
In vitro activity of natural and chemical products on sporulation of Eimeria species oocysts of chickens.
Gadelhaq Sahar M,Arafa Waleed M,Abolhadid Shawky M
This study was designed to investigate the ability of two herbal extracts and different chemical substances to inhibit or disrupt sporulation of Eimeria species oocysts of the chickens. The two herbal extracts were Allium sativum (garlic) and Moringa olifiera while the chemical substances included commercial disinfectants and diclazuril. Field isolates of Eimeria oocysts were propagated in chickens to obtain a continuous source of oocysts. The collected unsporulated oocysts (10oocysts/5 ml) were dispensed into 5 cm Petri dish. Three replicates were used for each treatment. The treated oocysts were incubated for 48 h at 25-29 °C and 80% relative humidity. The results showed that herbal extracts, the commercial recommended dose of Dettol, TH4, Phenol, VirkonS, and Diclazuril 20% have no effect on the sporulation. While Sodium hypochlorite showed a significant degree of sporulation inhibition reached to 49.67%. Moreover, 70% ethanol, and 10% formalin showed 100% sporulation inhibition. It was concluded that 70% ethanol and 10% formalin are the most effective methods to inhibit Eimeria species sporulation.
Anticoccidial effects of Galla rhois extract on Eimeria tenella-infected chicken.
Lee Hyun-A,Hong Sunhwa,Chung Yung-Ho,Song Ki-Duk,Kim Okjin
Laboratory animal research
Anticoccidial effects of Galla rhois (GR) extract were evaluated in chickens after oral infection with Eimeria tenella. This study was performed using 3-day-old chickens (n=30). The animals were divided into 3 groups as follows: GR 0.5%/infected (n=10), untreated/infected (n=10), and non-infected control (n=10). The chickens were fed a standard diet supplemented with or without GR for 1 week before infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of GR on E. tenella infection were assessed by 2 parameters, number of fecal oocysts and body weight gain, and the results of the polymerase chain reaction (PCR). The GR-fed chickens produced significantly lower number of fecal oocysts (P<0.05) than the E. tenella-infected chickens who were fed the standard diet. In addition, GR-based diet improved the loss of body weight caused by E. tenella infection. Positive findings of PCR were identified by distinct bands in the samples of E. tenella-inoculated chickens. However, PCR analysis revealed no E. tenella oocysts in the feces of GR-fed chickens. Our data showed that GR extracts had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of novel anticoccidial drugs.
Comparative effects of Herba Cox®, a commercial herbal extract, on rabbits (Oryctolagus cuniculus) experimentally infected with Eimeria stiedae.
Eladl Abdelfattah H,Mahgoub Hebatallah A,El-Shafei Reham A,Al-Kappany Yara M
Comparative immunology, microbiology and infectious diseases
The objective of this study was to evaluate the efficacy of Herba Cox®, a commercial herbal compound containing extracts from Bombax malabaricum, Aegle marmelos, Anethum foeniculum, Resina salvia, Ferula asafoetida and Papaver somniferum, for the treatment of rabbit hepatic coccidiosis. Thirty rabbits were allocated into three groups (10 × 3), the G1 group served as a negative control group, G2 group (positive control group) was infected with 5 × 10 sporulated E. stiedaeoocysts and served as infected-untreated group, and G3 group was infected with 5 × 10 sporulated E. stiedaeoocysts and treated with Herba Cox®, 1 ml/liter of drinking water, starting 7 days before infection and continuing for 4 weeks post-infection. When compared to the infected group (G2), body weight and weight gain were significantly (P ≤ 0.05) increased, the feed conversion rate was improved and no mortality was detected in infected treated group (G3) and similar to negative control group (G1). In addition, faecal oocyst output and liver enzymes were significantly decreased. Malondialdehyde, nitric oxide, and glutathione concentrations observed in G3 were similar to those in G1. In infected-untreated rabbits (G2), the haemoglobin, lymphocytes, and CD4+/ CD8+ ratio were significantly decreased, while the total leukocyte count, percentage of heterophils, and heterophil/lymphocyte ratio were increased. Significantly more severe histopathological hepatic lesions were observed in G2 when compared to G1 and G3. In conclusion, the obtained results showed that Herba Cox® should be considered a safe and novel effective compound for the treatment of E. stiedae infection in rabbits.
Anti-coccidial properties and mechanisms of an edible herb, Bidens pilosa, and its active compounds for coccidiosis.
Yang Wen-Chin,Yang Cheng-Ying,Liang Yu-Chuan,Yang Chu-Wen,Li Wei-Qun,Chung Chih-Yao,Yang Meng-Ting,Kuo Tien-Fen,Lin Chuen-Fu,Liang Chih-Lung,Chang Cicero Lee-Tian
Avian coccidiosis is an economically important disease in the poultry industry. In view of the disadvantages of anti-coccidial drugs in chickens, edible plants and their compounds are re-emerging as an alternative strategy to combat this disease. A previous publication reported that the edible plant B. pilosa showed promise for use against coccidiosis. Here, we first investigated into the anti-coccidial effects of B. pilosa. We found that B. pilosa at 100 ppm or more significantly suppressed E. tenella as evidenced by reduction in mortality rate, oocyst excretion and gut pathological severity in chickens and its minimum prophylactic duration was 3 days. Next, we explored the mode of action of anti-coccidial mechanism of B. pilosa. The E. tenella oocysts were not directly killed by B. pilosa; however, administration of the plant suppressed oocyst sporulation, sporozoite invasion, and schizonts in the life cycle of E. tenella. Besides, B. pilosa boosted T cell-mediated immunity. Finally, we characterized the related anti-coccidial phytochemicals and their mode of action. One of three potent polyynes present in B. pilsoa, Compound 1 (cytopiloyne), acted against coccidiosis in chickens in a similar manner to B. pilosa. These data illustrate the anti-coccidial potency and mechanism of B. pilosa and one of its active compounds, and provide a cornerstone for development of novel herbal remedies for avian coccidiosis.
Effects of pre-starter and starter diets varying in amino acid density given to broiler chickens that received coccidiosis vaccination at hatch1.
Cloft S E,Rochell S J,Macklin K S,Dozier W A
An experiment was conducted to evaluate broilers vaccinated against coccidiosis fed combinations of pre-starter and starter diets varying in digestible amino acid (dAA) density from 1 to 19 d of age on subsequent growth performance and meat yield. A total of 1,800 Yield Plus Ross × Ross 708 male broilers were allocated to 60 floor pens and assigned to 1 of the 6 treatments. Four pre-starter diets varying in dAA density [1.15% dLys, 1.25% dLys, 1.35% dLys, and Positive Control (1.15% dLys + Diclazuril)] were fed until 9 d of age. Then, 3 starter diets varying in dAA density [1.15% dLys, 1.25% dLys, and PC (1.15% dLys + Diclazuril)] were given from 10 to 19 d of age. All diets were formulated to similar dAA ratios to dLys; hence, resulting in increasing amino acid (AA) density among the dietary treatments. Birds were given common grower and finisher diets for the remainder of the experiment. At 19, 27, and 40 d of age, broilers that received the 1.35% and 1.25% dLys density pre-starter and 1.25% dLys density starter diets had increased BW gain (P ≤ 0.006) compared with broilers fed the 1.15% dLys density diets. Broilers that received the 1.15% dLys density pre-starter diet had a lower cumulative BW gain (P = 0.007) than those fed the 1.35% and 1.25% dLys density pre-starter diets. Broilers that received the 1.35% and 1.25% dLys density pre-starter and 1.25% dLys density starter diets had the heaviest (P ≤ 0.008) total breast weights. Increasing the pre-starter dAA density enhanced (P ≤ 0.03) total breast weight by 28 g (1.25% dLys density) and 51 g (1.35% dLys density). Results from this study indicated that increasing dAA density from 1 to 19 d of age enabled a 1 to 19 d dAA intake increase of 4.8 g, which enhanced cumulative growth and meat yield of broilers vaccinated against coccidiosis.
Effects of coccidiosis vaccination administered by in ovo injection on Ross 708 broiler performance through 14 days of post-hatch age.
Sokale A O,Zhai W,Pote L M,Williams C J,Peebles E D
Effects of the in ovo injection of a commercial coccidiosis vaccine on various hatching chick quality variables and 14 d post-hatch (dph) oocyst shedding have been previously examined. The current study was designed to examine the performance of Ross 708 broilers during the 14 dph period of oocyst shedding following the application of the coccidiosis vaccine. On each of 7 replicate tray levels of a single-stage incubator, a total of 4 treatment groups was randomly represented, with each treatment group containing 63 eggs. Treatments were administered using a commercial multi-egg injector on d 18.5 of incubation. The treatments included 3 control groups (non-injected, dry-punch, and diluent-injected) and one treatment group (injected with diluent containing Inovocox EM1 vaccine). On d 21 of incubation, 20 chicks from each of the 28 treatment-replicate groups were placed in corresponding wire-floored battery cages. Mortality, feed intake (FI), BW gain (BWG), and feed conversion ratio (FCR) were determined for the zero to 7, 7 to 14, and cumulative zero to 14 dph intervals. There were no significant treatment effects on mortality in any interval or on BW at zero dph. There were significant treatment effects on BW at 7 and 14 dph, on BWG and FI in the zero to 7, 7 to 14, and zero to 14 dph intervals, and on FCR in the 7 to 14 and zero to 14 dph intervals. Although the performance variables of birds belonging to the diluent-injected and vaccine-injected groups were not significantly different, the 14 dph BW, 7 to 14 dph FI, and zero to 14 dph BWG and FI of birds belonging to the vaccine treatment group were significantly higher than those in birds belonging to the non-injected control group. It was concluded that use of the Inovocox EM1 vaccine in commercial diluent has no detrimental effect on the overall post-hatch performance of broilers through 14 dph.
Pharmacokinetics and oral bioavailability of amoxicillin in chicken infected with caecal coccidiosis.
Journal of veterinary pharmacology and therapeutics
Chicken infected with caecal coccidiosis (Eimeria tenella) was used to evaluate the effect of coccidiosis on the pharmacokinetic and bioavailability of amoxicillin. The level of amoxicillin was estimated by high-performance chromatography (HPLC) to calculate the pharmacokinetic parameters and oral bioavailability. For i.v. injection of amoxicillin, Vd and CL were 0.29 and 0.27 (mg/kg)/(μg/mL)/h, respectively. Compared with healthy chicken, intravenous injection of amoxicillin in the infected chicken showed higher distribution and elimination constants, delayed clearance and statistically significant higher AUC and MRT. Oral administration in healthy chicken was accompanied by rapid absorption and high bioavailability with Tmax , Cmax and F about 1.03 h, 3.26 μg/mL and 40.2, respectively. Furthermore, oral administration in the infected chicken produced higher mean absorption time, delayed Tmax, lower Cmax, smaller AUC value and lower bioavailability (16.76). Based on these results, monitoring and adjustment of amoxicillin dosing could be practiced during the presence of coccidiosis. The measured Cmax values suggest the administration of 1.3-folds of the normal dose to maintain the normal maximal serum concentrations of amoxicillin in chicken infected with caecal coccidiosis.
Inclusion of Bacillus amyloliquefaciens strain TOA5001 in the diet of broilers suppresses the symptoms of coccidiosis by modulating intestinal microbiota.
Tsukahara Takamitsu,Inoue Ryo,Nakayama Keizo,Inatomi Takio
Animal science journal = Nihon chikusan Gakkaiho
Coccidiosis is an intestinal parasitic infection and one of the most prevalent and economically damaging diseases of chickens. Furthermore, coccidia-induced mucogenesis promotes secondary colonization by Clostridium perfringens, a major pathogen of chickens that causes necrotic enteritis. Our previous work found that supernatant of a culture of Bacillus amyloliquefaciens strain TOA5001 (BA) inhibited the growth of C. perfringens on Gifu anaerobic broth medium. Accordingly, we evaluated the effectiveness of dietary BA administration in inhibiting C. perfringens colonization of the intestine in broilers that were experimentally infected with coccidia. Ten healthy broilers from a BA-supplemented (2 × 10 colony-forming units/g of feed) broiler group and 10 from a non-treated group were challenged with Eimeria tenella and E. maxima (5000 oocysts of each species/chick) at 28 days old. At 36 days old, five chicks from each group were slaughtered, whereas the remaining five in each group were killed at 49 days old. Dietary BA administration into Eimeria-challenged birds reduced coccidial symptoms such as intestinal lesions. It also modified the cecal microbiota through suppressing C. perfringens and E. coli colonization, and inducing domination of Faecalibacterium prausnitzii, the Lactobacillus group and unknown Lachnospiraceae genera by bacterial DNA-based metagenome analyses. B. amyloliquefaciens TOA5001 supplementation suppressed the symptoms of coccidiosis by modulating cecal microbiota in Eimeria-challenged broilers.
Protective potential of diclazuril-treated oocysts against coccidiosis in layer chicks.
El-Ashram Saeed,Aboelhadid Shawky M,Arafa Waleed M,Gadelhaq Sahar M,Abdel-Razik Abdel-Razik H
Diclazuril, which is widely used for the prevention of coccidiosis in chickens, has a lethal effect on asexual and sexual stages of Eimeria spp. However, little is known about its effect on the exogenous stages of Eimeria spp. In this study, we evaluated the effect of in vitro treatment with 0.2% diclazuril on unsporulated and sporulated oocysts of Eimeria spp. For this purpose, a total of 180 male layer chicks aged one day were randomly divided into 5 experimental groups. Each group was divided into 3 replicates of 12 chicks each. Group 1 (G1) and Group 2 (G2) were negative (non-immunized and non-challenged) and positive (non-immunized and challenged) controls, respectively. Group 3 (G3) was immunized per os with 1.0 × 10 non-diclazuril treated-sporulated oocysts. Groups 4 (G4) was immunized per os with 0.2% diclazuril treated-unsporulated oocysts (1.0 × 10) in which diclazzuril didn't affect sporulation. Group 5 (G5) was immunized per os with 0.2% diclazuril treated-sporulated oocysts (1.0 × 10). Chicks of G2, G3, G4, and G5 were challenged with 7.5 × 10 untreated sporulated oocysts at the age of 21 days, while the group 1 chicks remained unchallenged. G4 and G5 animals immunized with 0.2% diclazuril-treated oocysts showed a significant decrease in bloody diarrhea severity, lesion scores, and oocyst counts in comparison to those immunized with untreated oocysts. Furthermore, histopathologic findings showed a low number of parasitic stages in cecal tissues in G4 and G5. A significant increased body weight gain was observed in Gs 4 and 5 in comparison to G2. In addition, expression levels of IL-2, IL-12, and IFN-γ were significantly increased in G4 and G5. In conclusion, diclazuril is effective in attenuating Eimeria oocysts and thus provides an alternative approach for using diclazuril-treated oocysts to protect chicks against Eimeria challenge.
Control of eight predominant Eimeria spp. involved in economic coccidiosis of broiler chicken by a chemically characterized essential oil.
Barbour E K,Bragg R R,Karrouf G,Iyer A,Azhar E,Harakeh S,Kumosani T
Journal of applied microbiology
AIM:To control eight most predominant Eimeria spp. involved in the economic disease of coccidiosis in broiler chicken, by a chemically characterized essential oil of eucalyptus and peppermint. METHODS AND RESULTS:The experimental design consisted of 160 day-old-broiler chicks, divided into four equal groups (G1 , G2 , G3 and G4 ), with 40 birds per group. Each group was divided into four equal subgroups. Birds in G1 were deprived of essential oil treatment and of Eimeria challenge. Birds in G2 were unchallenged, and administered the essential oil in drinking water at 0.69 ml kg(-1) body weight. Birds in G3 were untreated with essential oil, and each of its four subgroups was challenged at a different age (14, 21, 28 and 35 days). Birds in G4 were treated with essential oil, and challenged in the same manner as for G3 . Equal number of birds from all subgroups (n = 10) were sacrificed at the sixth day after the time allocated for each challenge. The 6 day incubation period post challenge resulted in respective mean per cent weight increase in G2 and G1 birds equivalent to 57.8 and 53.1% (P < 0.05). In addition, the essential oil improved the per cent weight increase in challenged birds (54.6%) compared to the challenged-untreated birds (18.6%) (P < 0.05). The mean feed conversion, mortality, intestinal lesion scores and oocyst counts were significantly reduced in the challenged-treated birds compared to the challenged-untreated birds (P < 0.05). CONCLUSIONS:The results support the hypothesis of using the essential oils of eucalyptus and peppermint to control the most prevalent Eimeria spp. involved in coccidiosis of broiler chicken, helping in improvement of their production, alleviation of lesions and reduction in intestinal oocyst counts. SIGNIFICANCE AND IMPACT OF THE STUDY:This study provides information about the possibility of using this blend of essential oil as a coccidiostat for the protection of broiler chickens against the prevalent eight Eimeria spp. of coccidiosis.
Dietary oregano essential oil alleviates experimentally induced coccidiosis in broilers.
Mohiti-Asli M,Ghanaatparast-Rashti M
Preventive veterinary medicine
An experiment was conducted to determine the effects of oregano essential oil on growth performance and coccidiosis prevention in mild challenged broilers. A total of 250 1-d-old chicks were used in a completely randomized design with 5 treatments and 5 replicates with 10 birds in each replication. Experimental treatments included: (1) negative control (NC; unchallenged), (2) positive control (PC; challenged with sporulated oocysts of Eimeria), (3) PC fed 200 ppm Diclazuril in diet, (4) PC fed 300 ppm oregano oil in diet, and (5) PC fed 500 ppm oregano oil in diet. At 22 d of age, all the experimental groups except for NC were challenged with 50-fold dose of Livacox T as a trivalent live attenuated coccidiosis vaccine. On d 28, two birds were slaughtered and intestinal coccidiosis lesions were scored 0-4. Moreover, dropping was scored in the scale of 0-3, and oocysts per gram feces (OPG) were measured. Oregano oil at either supplementation rate increased body weight gain (P=0.039) and improved feed conversion ratio (P=0.010) from d 22 to 28, when compared with PC group. Using 500 ppm oregano oil in challenged broilers diet increased European efficiency factor than PC group (P=0.020). Moreover, challenged broilers fed 500 ppm oregano oil or Diclazuril in diets displayed lower coccidiosis lesions scores in upper (P=0.003) and middle (P=0.018) regions of intestine than PC group, with the effect being similar to unchallenged birds. In general, challenged birds fed 500 ppm oregano oil or Diclazuril in diets had lower OPG (P=0.001), dropping scores (P=0.001), litter scores (P=0.001), and pH of litter (P=0.001) than PC group. It could be concluded that supplementation of oregano oil at the dose of 500 ppm in diet may have beneficial effect on prevention of coccidiosis in broilers.
Use of discriminant analysis for the evaluation of coccidiosis resistance parameters in chickens raised in hot humid tropical environment.
Adenaike A S,Peters S O,Adeleke M A,Fafiolu A O,Takeet M I,Ikeobi C O N
Tropical animal health and production
Coccidiosis endemicity remains a major challenge in poultry production in the tropics and all over the world. In order to develop predictive tool for identification of chickens that are at risk of coccidiosis among Nigerian indigenous chickens, body weight gain (BWG) and hematological variables were determined for chickens infected with Eimeria tenella (female = 60, male = 63) and uninfected (female = 51, male = 45). The hematological variables analyzed include the following: packed cell volume (PCV, %), white blood cells (WBC, × 10/μl), and red blood cells (RBC, × 10/μl), as well as differential leucocyte percentages of neutrophils, lymphocytes, monocytes, and eosinophils. Body weight gain was determined at days 3, 6, 9, 12, and 15. Of the 12 variables analyzed, BWG at day 3, monocyte, PCV, and WBC in males and BWG at days 6, 9, and 12, PCV, and WBC in female chickens showed significant (P ≤ 0.01) difference between the infected and uninfected. Stepwise discriminant analysis evolved a model that could distinguish uninfected from Eimeria-infected chickens. Packed cell volume, WBC, BWG at day 3, and lymphocytes emerged the most discriminant between uninfected and Eimeria-infected chickens in male chickens. In female chickens, PCV, RBC, and BWG at day 3 were identified as most discriminant variables in separating the uninfected from Eimeria-infected chickens. Therefore, this study suggests that routine blood test and estimates of body weight gain could serve as a useful tool for identifying chickens that may be at risk of coccidiosis, enabling improvement of preventive measures.
Serological diagnosis of Toxoplasmosis disease using a fluorescent immunosensor with chitosan-ZnO-nanoparticles.
Medawar-Aguilar Victoria,Jofre Claudio F,Fernández-Baldo Martín A,Alonso Andrés,Angel Sergio,Raba Julio,Pereira Sirley V,Messina Germán A
This article describes a microfluidic LIF immunosensor for the quantitative determination of anti-Toxoplasma gondii IgG (anti-T. gondii) specific antibodies. The serological detection of these antibodies plays a crucial role in the clinical diagnosis of toxoplasmosis. Zinc oxide nanoparticles (ZnO-NPs) obtained by wet chemical procedure were covered with chitosan and then used to conjugate T-gondii antigens into the central microfluidic channel. Serum samples containing anti-T-gondii IgG antibodies were injected into the immunosensor where they interact immunologically with T. gondii antigens. Bound antibodies were quantified by the addition of anti-IgG antibodies labeled whit alkaline phosphatase (ALP). ALP enzymatically converts the non-fluorescent 4-methylumbelliferyl phosphate (4-MUP) to soluble fluorescent methylumbelliferone that was measured using excitation at 355 nm and emission at 440 nm. The relative fluorescent response of methylumbelliferone is proportional to the concentration of anti-T. gondii IgG antibodies. The coefficients of variation are less than 4.73% for within-day assays and less than 6.34% for between-day assays. Results acquired by LIF immunosensor agree with those obtained by enzyme-linked immunosorbent assay method, suggesting that the designed sensor represents a promising tool for the quantitative determination of anti-T. gondii IgG antibodies of clinical samples.
Chitosan and silver nanoparticles: promising anti-toxoplasma agents.
Gaafar M R,Mady R F,Diab R G,Shalaby Th I
Toxoplasmosis is a worldwide infection caused by obligate intracellular protozoan parasite which is Toxoplasma gondii. Chitosan and silver nanoparticles were synthesized to be evaluated singly or combined for their anti-toxoplasma effects as prophylaxis and as treatment in the experimental animals. Results were assessed through studying the parasite density and the ultrastructural parasite changes, and estimation of serum gamma interferon. Weight of tissue silver was assessed in different organs. Results showed that silver nanoparticles used singly or combined with chitosan have promising anti-toxoplasma potentials. The animals that received these compounds showed statistically significant decrease in the mean number of the parasite count in the liver and the spleen, when compared to the corresponding control group. Light microscopic examination of the peritoneal exudates of animals receiving these compounds showed stoppage of movement and deformity in shape of the tachyzoites, whereas, by scanning electron microscope, the organisms were mutilated. Moreover, gamma interferon was increased in the serum of animals receiving these compounds. All values of silver detected in different tissues were within the safe range. Thus, these nanoparticles proved their effectiveness against the experimental Toxoplasma infection.
Inhibitory activity of chitosan nanoparticles against Cryptosporidium parvum oocysts.
Ahmed Shahira A,El-Mahallawy Heba S,Karanis Panagiotis
Cryptosporidium is a ubiquitous harsh protozoan parasite that resists many disinfectants. It remains viable and infective for a long time in water and food causing global outbreaks. Chitosan (the deacetylated chitin molecule) was used in its nanosuspension form to evaluate its effect against Cryptosporidium parvum. The experiments were performed in vitro in serial concentrations and confirmed in mice in vivo infectivity assay. Chitosan nanoparticles (Cs NPs) were toxic to Cryptosporidium oocysts. The effect appeared to decrease the number of Cryptosporidium oocysts and altered their content. The destruction rate of oocysts was dependent on the dose of chitosan and the time of exposure (P < 0.05). Higher doses of Cs NPs over a prolonged period exhibited a significantly higher destruction rate. Using staining and light microscopy, remarkable destructive changes were observed in the oocysts' morphology. The minimal lethal dose for > 90% of oocysts was 3000 μg/ml, no mice infections in vivo were observed. The results in this study elucidate Cs NPs as an effective anti-cryptosporidial agent.
Anticoccidial activities of Chitosan on Eimeria papillata-infected mice.
Abdel-Latif Mahmoud,Abdel-Haleem Heba M,Abdel-Baki Abdel-Azeem S
Eimeria spp. multiply within the intestinal tract causing severe inflammatory responses. Chitosan (CS), meanwhile, has been shown to exhibit anti-inflammatory activities in different experimental models. Here, we investigated the effect of CS on the outcome of inflammation caused by Eimeria papillata in the mouse intestine. Investigations were undertaken into the oocyst output in feces and developmental stages and goblet cells in intestinal tissue. Assays for lipid peroxidation, nitric oxide (NO), and myeloperoxidase (MPO) were also performed. T cells in intestinal tissue were counted using immunohistochemistry while total IgA in serum or intestinal wash was assayed using ELISA. In addition, mRNA expression of tumor necrosis factor alpha (TNF-α), transforming growth factor β (TGF-β), interleukin (IL)-10, and IL-4 were detected using real-time PCR. The data indicated a reduction in both oocyst output and in the number of parasite developmental stages following CS treatment, while the goblet cell hypoplasia in infected mice was also inhibited. CS decreased lipid peroxidation, NO, and MPO but did not alter the T cell count or IgA levels in comparison to the infected group. The expression of TNF-α and TGF-β decreased but IL-10 and IL-4 increased after CS treatment in comparison to the non-treated infected group. In conclusion, CS showed anti-inflammatory and protective effects against E. papillata infection.