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.