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1.
Avian Dis ; 65(1): 77-85, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-34339126

RESUMO

Infection of poultry with Eimeria spp., the causative agent of coccidiosis, can predispose birds to necrotic enteritis (NE) caused by netB gene-positive strains of Clostridium perfringens. The detection of Eimeria spp., C. perfringens, and netB were examined in settled dust from broiler flocks under experimental and field conditions. Dust samples were collected from settle plates twice weekly from two experimental flocks inoculated with three species of pathogenic Eimeria in 9-day-old chicks, followed by netB gene-positive C. perfringens 5 days later to produce subclinical and clinical NE. A noninoculated flock was sampled weekly from day 0 and served as a control flock. An additional 227 dust samples from commercial broiler flocks were collected at the end-of-batch (6-7 wk of age; one scraped dust sample per flock). In the NE-subclinical and NE-clinical flocks, high levels of Eimeria spp. and C. perfringens were detected after inoculation followed by a gradual decline over time. In the control flock, C. perfringens and netB were detected at low levels. No significant effect of sampling location was evident on Eimeria spp., C. perfringens, and netB load within poultry houses. These results provide evidence that Eimeria spp., C. perfringens, and netB gene copies can be readily measured in poultry dust samples collected in settle plates and may provide an alternative sampling method for monitoring flock coccidiosis and NE status. Further studies are required to assess the utility for such a test in commercial flocks.


Assuntos
Toxinas Bacterianas/análise , Galinhas , Infecções por Clostridium/veterinária , Clostridium perfringens/isolamento & purificação , Coccidiose/veterinária , Eimeria/isolamento & purificação , Enterotoxinas/análise , Doenças das Aves Domésticas/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/microbiologia , Coccidiose/epidemiologia , Coccidiose/parasitologia , Poeira , New South Wales/epidemiologia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/parasitologia
2.
Poult Sci ; 100(9): 101369, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34333388

RESUMO

Compounds in microalgae-derived feed ingredients in poultry diets may improve intestinal physiology and immunity to protect against damage induced by physiological and pathogen challenges, but mechanisms are examined sparingly. The study objective was to evaluate changes to intestinal morphology, permeability, and systemic immunity in broilers fed a proprietary microalgae ingredient during 2 separate challenge studies. In study 1, two replicate 28 d battery cage trials used 200 Ross 308 broilers each (n = 400) fed a control diet ± 0.175% algae ingredient. Half of the birds were subjected to a 12 h feed restriction challenge and fluorescein isothiocyanate dextran (FITC-D) intestinal permeability assay on d 28. Study 2 used 800 broilers randomly assigned to the same dietary treatments and housed in floor pens for 42 d. At d 14, intestine and spleen samples were collected from 10 birds/ diet. Half of the remainder was orally inoculated with 10X Coccivac-B52 vaccine in a 2 × 2 factorial treatment design (diet and Eimeria inoculation). The FITC-D assay was conducted at 1, 3, 7, and 14 d post-inoculation (pi) while intestinal and spleen samples were collected at 3, 7, 14, and 28 dpi for histomorphology and flow cytometric immune cell assessment. Study 1 validated intestinal leakage via FITC-D absorbance induced by feed restriction but showed no algae-associated protective effects. In study 2, algae preserved intestinal integrity during coccidiosis (P = 0.04) and simultaneously protected jejunal villus height as early as 7dpi (P < 0.0001), whereas intestinal damage resolution in control birds did not occur until 14 dpi. Algae inclusion increased splenic T cells in unchallenged broilers at d 14 by 29.6% vs. control (P < 0.0001), specifically γδ T cell populations, without impacting performance (P < 0.03). During Eimeria challenge, splenic T cells in algae-fed birds did not show evidence of recruitment to peripheral tissues, while control birds showed a 16.7% reduction compared to their uninoculated counterparts from 3 to 7 dpi (P < 0.0001). This evidence suggests the algae ingredient altered the immune response in a manner that reduced recruitment from secondary lymphoid organs in addition to protecting intestinal physiology.


Assuntos
Coccidiose , Eimeria , Doenças das Aves Domésticas , Ração Animal/análise , Animais , Galinhas , Coccidiose/prevenção & controle , Coccidiose/veterinária , Dieta/veterinária , Suplementos Nutricionais , Imunidade , Intestinos
3.
Avian Dis ; 65(1): 132-137, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-34339132

RESUMO

Necrotic enteritis (NE) is a common disease that causes great economic loss to the broiler industry due to mortality and reduced performance. Although Clostridium perfringens (CP) is a necessary component of this disease, coccidia species are a well-defined predisposing factor that exacerbates the condition. Different Eimeria species have been reported to influence NE to different degrees. In a pair of experiments, six different Eimeria species were evaluated in the presence and absence of C. perfringens. Male broiler chicks were housed in battery cages for the duration of both experiments. Feed conversion, body weight gain, and NE mortality were reported in both experiments. Experiment 1 challenged birds with E. maxima, E. acervulina, E. tenella, E. necatrix, and E. brunetti at day 13 and subsequently inoculated birds with CP on days 18, 19, and 20. In the second experiment, E. maxima, E. acervulina, E. tenella, and E. praecox were inoculated on day 15 and challenged with CP on days 17, 18, 19, 20, 21, and 22 of the experiment. In the first experiment, E. acervulina, E. brunetti, E. maxima, and E. necatrix with the addition of CP all stimulated necrotic enteritis mortality. In the second experiment, E. praecox had minimal impact on performance during the challenge (14-23 days) while E. maxima + CP decreased body weight gain and increased mortality compared to the CP alone control. Eimeria maxima had the highest mortality (21.9%) in this experiment followed by E. acervulina (6.3%). The remaining Eimeria with added CP in the second experiment did not induce NE mortality. While the challenge with CP alone did not induce mortality, feed conversion was increased compared to the unchallenged control group. When using isolated Eimeria species in these experiments, disturbances created by E. brunetti and E. maxima resulted in the most-severe challenges. These experiments highlight the NE risk of these species of Eimeria and give insight into how other species interact with the host in a controlled CP challenge model.


Assuntos
Galinhas , Infecções por Clostridium/veterinária , Coccidiose/veterinária , Eimeria/fisiologia , Enterite/veterinária , Necrose/veterinária , Doenças das Aves Domésticas , Animais , Infecções por Clostridium/microbiologia , Clostridium perfringens/fisiologia , Coccidiose/parasitologia , Enterite/microbiologia , Enterite/parasitologia , Masculino , Necrose/microbiologia , Necrose/parasitologia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/parasitologia , Distribuição Aleatória , Especificidade da Espécie
4.
Avian Dis ; 65(1): 149-158, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-34339134

RESUMO

With growing cross-disciplinary collaboration among researchers, it is increasingly important to record detailed methodology to prevent the repetition of preliminary experiments. The purpose of this paper is to explain the development of a coccidiosis challenge model for the investigation of dietary interventions to coccidiosis in broiler chickens. The objectives are to select a dose of mixed species coccidial vaccine and evaluate the suitability (ability to produce a consistent, marked change) of selected response variables important to nutritional studies at different times postinfection (PI). Coccivac-B and Coccivac-B52 (Merck Animal Health) were evaluated as the source of coccidia in three trials. Trials 1 and 2 were randomized complete block designs with four doses (0, 10, 20, or 30 times (×) label dose) of Coccivac-B administered to 12 replicate cages of six birds by repeater pipette (Trial 1) or gavaging needle (Trial 2). Trial 3 used a completely randomized design with 0× or 30× label dose of Coccivac-B52 administered by gavaging needle to six replicate cages of six birds. Birds were gavaged at 15 days of age, and response criteria were evaluated 7 days PI in all trials and again at 10 days PI in Trials 1 and 2. All means are reported in order of increasing coccidia dose with significance accepted at P ≤ 0.05. Broiler performance was not affected by coccidia in Trials 1 or 3 but grew poorer with increasing dose from 0 to 7 days PI in Trial 2 (body weight gain, 465, 421, 388, 365 g; feed to gain, 1.37, 1.47, 1.52, 1.58). As coccidia dose increased, nitrogen corrected apparent metabolizable energy decreased (Trial 1, 3387, 3318, 3267, 3170 kcal kg-1; Trial 2, 3358, 2535, 2422, 2309 kcal kg-1; Trial 3, not measured), while relative weight, length, and content for intestinal sections increased (Trials 1through 3). Gross lesion (duodenum, jejunum/ileum, ceca) and oocyst count scores (jejunum/ileum, ceca) increased with dose; however, gross scoring often suggested infection in unchallenged birds, a finding unsupported by oocyst count scores. At 7 days PI there was no correlation between midgut gross lesion score and midgut oocyst count score (r = 0.06, P = 0.705), but cecal scores were weakly correlated (r = 0.55, P < 0.001). Administering coccidia via repeater pipette (Trial 1) resulted in respiratory distress in some birds, while use of the gavaging needle (Trials 2 and 3) successfully induced intestinal damage in chickens without resulting in coccidia related mortality. Thirty times the label dose at 7 days PI resulted in the greatest number of response variables that produced a consistent, marked change. Therefore, consideration should be given to these conditions when designing future coccidiosis challenge models using vaccines as a source of coccidia.


Assuntos
Galinhas , Coccidiose/veterinária , Eimeria/imunologia , Doenças das Aves Domésticas/prevenção & controle , Vacinas Protozoárias/administração & dosagem , Ração Animal/análise , Animais , Coccidiose/parasitologia , Coccidiose/prevenção & controle , Dieta/veterinária , Suplementos Nutricionais , Masculino , Oocistos , Doenças das Aves Domésticas/parasitologia , Vacinas Atenuadas/administração & dosagem
5.
Poult Sci ; 100(10): 101391, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34428644

RESUMO

A series of 6 floor pen trials was conducted to determine the effects of a quillaja and yucca combination product on the performance and carcass traits of growing broiler chickens vaccinated for coccidiosis at the hatchery. In each of the trials graded levels (0, 250, and 500 ppm) of a quillaja and yucca combination (QY) were fed to Ross 708 broilers for the duration of each 42 d test. Trials were arranged in completely randomized block designs involving a minimum of 11 blocks per trial. At the start of each trial, pens contained 55 broilers. In order to provide each bird with an enteric disease challenge, 5 kg commercial broiler litter containing 104 CFU Clostridium perfringens per gram was placed in each pen. In addition, the sporulated oocysts of Eimeria acervulina and E. maxima were added to each pen at the outset of each test. At d 21 of the trials, coccidial lesion scores, mortality and performance were determined; final performance and total mortality were assessed at 42 d. At the completion of each test, 10 birds of average body weight per pen were selected for carcass evaluations; whole and chilled carcass yield were determined, and pre- and post-chill breast measurements were made. A combined analysis of the results of the 6 trials (75 replications per treatment) was used to determine treatment effects and each variable was assessed by linear regression analysis. Results indicated that QY significantly reduced mortality and coccidial lesions scores at d 21 (P < 0.05). Performance was significantly improved by both levels of QY at 21 and 42 d, and significant linear effects were observed for these variables (P < 0.05). All carcass characteristics were significantly improved by QY administration and significant linear responses were observed for each carcass variable (P < 0.05). These results indicate that by reducing intestinal disease challenge, QY provided linear improvements in performance. In addition, QY positively affected carcass parameters as each variable responded linearly to QY feeding (P < 0.05).


Assuntos
Coccidiose , Eimeria , Doenças das Aves Domésticas , Yucca , Ração Animal/análise , Animais , Galinhas , Coccidiose/prevenção & controle , Coccidiose/veterinária , Dieta , Esterco , Doenças das Aves Domésticas/prevenção & controle , Quillaja
6.
Front Cell Infect Microbiol ; 11: 703413, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34336724

RESUMO

Eimeria bovis is an intracellular apicomplexan parasite that causes considerable economic losses in the cattle industry worldwide. During the first merogony, E. bovis forms large macromeronts with >140,000 merozoites I in host endothelial cells. Because this is a high-energy demanding process, E. bovis exploits the host cellular metabolism to fulfill its metabolic requirements. We here analyzed the carbohydrate-related energetic metabolism of E. bovis-infected primary bovine umbilical vein endothelial cells during first merogony and showed that during the infection, E. bovis-infected culture presented considerable changes in metabolic signatures, glycolytic, and mitochondrial responses. Thus, an increase in both oxygen consumption rates (OCR) and extracellular acidification rates (ECAR) were found in E. bovis-infected host cells indicating a shift from quiescent to energetic cell status. Enhanced levels of glucose and pyruvate consumption in addition to increased lactate production, suggesting an important role of glycolysis in E. bovis-infected culture from 12 days p.i. onward. This was also tested by glycolytic inhibitors (2-DG) treatment, which reduced the macromeront development and diminished merozoite I production. As an interesting finding, we observed that 2-DG treatment boosted sporozoite egress. Referring to mitochondrial activities, intracellular ROS production was increased toward the end of merogony, and mitochondrial potential was enhanced from 12 d p. i. onward in E. bovis-infected culture. Besides, morphological alterations of membrane potential signals also indicated mitochondrial dysfunction in macromeront-carrying host endothelial culture.


Assuntos
Eimeria , Animais , Bovinos , Células Endoteliais/metabolismo , Glicólise , Merozoítos , Mitocôndrias
7.
Parasite ; 28: 61, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34374643

RESUMO

Eimeria spp. cause the disease coccidiosis, which results in chronic wasting of livestock and can lead to the death of the animal. The disease, common worldwide, has caused huge economic losses to the cattle industry in particular. This is the first systematic review and meta-analysis of the prevalence of bovine Eimeria in China. Our search of five databases including PubMed, ScienceDirect, China National Knowledge Infrastructure (CNKI), Chongqing VIP, and Wan Fang for articles published up to February 29, 2020 on the prevalence of Eimeria in cattle in mainland China yielded 46 articles, in which the prevalence of cattle ranged from 4.6% to 87.5%. The rate of bovine Eimeria infection has been decreasing year by year, from 57.9% before 2000 to 25.0% after 2015, but it is still high. We also analyzed the region, sampling years, detection methods, feeding model, seasons, and species of bovine Eimeria. We recommend that prevention strategies should focus on strengthening detection of Eimeria in calves in the intensive farming model.


Assuntos
Doenças dos Bovinos , Coccidiose , Eimeria , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , China/epidemiologia , Coccidiose/epidemiologia , Coccidiose/veterinária , Fezes , Prevalência , Fatores de Risco
8.
Poult Sci ; 100(9): 101329, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34333387

RESUMO

This study evaluated growth performance and cross-protection against Eimeria spp. using a subunit coccidia vaccine in 2 independent challenge experiments. In both trials, chickens were challenged with E. acervulina, E. maxima, and E. tenella oocysts. In Exp 1, 1000-day-old chickens were allocated in one of 2 treatments 1) Control group; 2) Biotech Vac Cox group. The vaccine was orally gavaged on d 2 and 16 of life and coccidia challenge was on d 21. Performance parameters were evaluated on d 21, 35, and 42. On d 34, coccidia lesions were scored. Oocysts per gram of feces (OPG) were evaluated on d 28, 35, and 42. In Exp 2, 900-day-old chickens were assigned in one of 2 treatments 1) Control group; 2) Biotech Vac Cox group. The vaccine was orally gavaged on d 2 and 16 of life and coccidia challenge was on d 21. Performance parameters were evaluated on d 21, 27, 35, and 42, and lesion scores and OPG at d 27. In Exp 1, chickens vaccinated had significantly lower feed intake (FI) at d 21 and feed conversion ratio (FCR) at d 35 compared to control chickens (P < 0.05). Vaccinated chickens showed a significant reduction (P ≤ 0.05) in OPG for E. maxima to nondetectable levels and for all coccidian species at d 42 compared to control chickens. In Exp 2, the chickens vaccinated showed a significant increase in BW, BW gain (BWG) and reduction in FCR on d 27, 35, and 42 (P ≤ 0.05). Vaccinated chickens had significantly lower (P ≤ 0.05) lesion scores for all 3 Eimeria species. Moreover, vaccinated chickens had a reduction in total OPG of 35.50% (P = 0.0739). Studies to evaluate the serological and mucosal immune response are currently being evaluated. This inactivated, orally delivered subunit vaccine offers significant cross-protection to Eimeria spp. and eliminates the needs to treat broilers with live oocysts, enhanced ease of use, and greater biosecurity to producers.


Assuntos
Coccidiose , Eimeria tenella , Eimeria , Doenças das Aves Domésticas , Ração Animal/análise , Animais , Biotecnologia , Galinhas , Coccidiose/prevenção & controle , Coccidiose/veterinária , Doenças das Aves Domésticas/prevenção & controle , Vacinas de Subunidades
9.
Parasitol Res ; 120(8): 2973-2979, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34236530

RESUMO

We detected Eimeria oocysts from Japanese green pheasants (Phasianus versicolor) at a zoo in Osaka, Japan. The oocyst isolates were subspherical or ovoidal shaped and measured 17.2 (range 14.7-20.0) µm in length and 14.8 (13.3-16.7) µm in width with a length/width (L/W) ratio of 1.2 (1.0-1.4) and each had one polar granule. The oocysts lacked a residuum and micropyle. Sporocysts measured 9.8 (6.7-13.3) µm in length and 5.9 (4.7-7.3) µm in width, with a L/W ratio of 1.2 (1.1-1.4). Compared to previously published values, this strain shows morphological similarities with an isolate of E. teetartooimia from ring-necked pheasants from other countries. Phylogenetic analysis of the 18S rRNA and mitochondrial cytochrome c oxidase subunit I genes places the isolate in a clade related to chicken Eimeria spp., such as E. acervulina or E. brunetti. Although further analysis is needed, this information can be helpful for the diagnosis and determination of virulence of Eimeria spp. in pheasants.


Assuntos
Coccidiose , Eimeria , Galliformes , Oocistos , Animais , Coccidiose/veterinária , Eimeria/citologia , Eimeria/genética , Fezes , Galliformes/parasitologia , Japão , Oocistos/citologia , Oocistos/genética , Filogenia
10.
Poult Sci ; 100(8): 101239, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34214749

RESUMO

Avian coccidiosis continues to be one of the costliest diseases of commercial poultry. Understanding the epidemiology of Eimeria species in poultry flocks and the resistance profile to common anticoccidials is important to design effective disease prevention and control strategies. This study examined litter samples to estimate the prevalence and distribution of Eimeria species among broiler farms in 4 geographic regions of Colombia. A total of 245 litter samples were collected from 194 broiler farms across representative regions of poultry production between March and August 2019. The litter samples were processed for oocysts enumeration and speciation after sporulation. End-point polymerase chain reaction (PCR) analysis was conducted to confirm the presence of Eimeria species. Anticoccidial sensitivity was determined with 160 Ross AP males in 5 treatment groups: noninfected, nonmedicated control (NNC), infected, nonmedicated control (INC), infected salinomycin treated (SAL, dose: 66 ppm), infected diclazuril treated (DIC, dose: 1 ppm), and infected methylbenzocuate-Clopidol treated (MET.CLO, dose: 100 ppm), All birds were orally inoculated with 1 × 106 sporulated oocysts using a 1 mL syringe, except for the NNC- group who received 1ml of water.Eimeria spp. were found in 236 (96.3%) out of 245 individual houses, representing 180 (92.8%) out of 194 farms. Eimeria acervulina was the most prevalent species (35.0%) followed by Eimeria tenella (30.9%), Eimeria maxima (20.4%), and other Eimeria spp. (13.6%). However, mixed species infections were common, with the most prevalent combination being mixtures of E. acervulina, E. maxima, E. tenella, and other species in 31.4% of the Eimeria-positive samples. PCR analysis identified E. acervulina, E. maxima, E. tenella, Eimeria necatrix, Eimeria mitis, and Eimeria praecox with variable prevalence across farms and regions. Anticoccidial sensitivity testing of strains of Eimeria isolated from 1 region, no treatment difference (P > 0.05) was observed in final weight (BW), weight gain (BWG) or feed conversion (FCR). For the global resistance index (GI) classified SAL and MET.CLO as good efficacy (85.79 and 85.49, respectively) and DIC as limited efficacy (74.52%). These results demonstrate the ubiquitous nature of Eimeria spp. and identifies the current state of sensitivity to commonly used anticoccidials in a region of poultry importance for Colombia.


Assuntos
Coccidiose , Coccidiostáticos , Eimeria , Doenças das Aves Domésticas , Animais , Galinhas , Coccidiose/tratamento farmacológico , Coccidiose/epidemiologia , Coccidiose/veterinária , Coccidiostáticos/uso terapêutico , Colômbia/epidemiologia , Fazendas , Masculino , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/epidemiologia
11.
Vet Res ; 52(1): 81, 2021 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-34108017

RESUMO

This study was conducted to investigate the impacts of graded severity of Eimeria maxima infection on the growth performance and intestine health of broiler chickens. Four different levels of E. maxima-challenged treatments were used, including a non-challenged control group, a low challenge (12 500 oocysts), a medium challenge (25 000 oocysts), and a high challenge dose (50 000 oocysts). There were eight replicate cages per treatment, with 12 birds in each cage, and chickens in the challenged groups orally received sporulated oocysts on day 14. Gastrointestinal permeability was measured by fluorescein isothiocyanate dextran at 5 days post-infection (dpi), whereas intestinal morphology and gene expression of nutrient transporters and tight junction proteins were determined at 6 dpi. The results demonstrate a linear reduction in growth performance, jejunal villus height, and jejunal integrity with graded challenge doses of E. maxima (P < 0.01). Moreover, linear regulation of nutrient transporters and tight junction proteins was a consequence of increasing Eimeria infection levels (P < 0.01). The linear increase of Claudin 1, cationic amino acid transporter, glucose transporter 1, and L-type amino acid transporter genes was associated with increased severity of coccidiosis (P < 0.01). Furthermore, expression of nutrient transporters located at the brush border membrane were down-regulated (P < 0.01) with increasing E. maxima inoculation dose. In conclusion, growth performance and key intestinal integrity biomarkers in broiler chickens were adversely influenced in a dose-dependent manner by E. maxima infection.


Assuntos
Galinhas , Coccidiose/veterinária , Eimeria/fisiologia , Expressão Gênica , Nutrientes/metabolismo , Doenças das Aves Domésticas/parasitologia , Animais , Transporte Biológico , Biomarcadores , Galinhas/genética , Galinhas/crescimento & desenvolvimento , Coccidiose/parasitologia , Intestinos/fisiologia , Masculino
12.
Parasit Vectors ; 14(1): 308, 2021 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-34099031

RESUMO

BACKGROUND: Coccidiosis caused by Eimeria stiedae is a widespread and economically significant disease of rabbits. The lack of studies on the life-cycle development and host interactions of E. stiedae at the molecular level has hampered our understanding of its pathogenesis. METHODS: In this study, we present a comprehensive transcriptome landscape of E. stiedae to illustrate its dynamic development from unsporulated oocysts to sporulated oocysts, merozoites, and gametocytes, and to identify genes related to parasite-host interactions during parasitism using combined PacBio single-molecule real-time and Illumina RNA sequencing followed by bioinformatics analysis and qRT-PCR validation. RESULTS: In total, 12,582 non-redundant full-length transcripts were generated with an average length of 1808 bp from the life-cycle stages of E. stiedae. Pairwise comparisons between stages revealed 8775 differentially expressed genes (DEGs) showing highly significant description changes, which compiled a snapshot of the mechanisms underlining asexual and sexual biology of E. stiedae including oocyst sporulation between unsporulated and sporulated oocysts; merozoite replication between sporulated oocysts and merozoites; and gametophyte development and gamete generation between merozoites and gametocytes. Further, 248 DEGs were grouped into nine series clusters and five groups by expression patterns, and showed that parasite-host interaction-related genes predominated in merozoites and gametocytes and were mostly involved in steroid biosynthesis and lipid metabolism and carboxylic acid. Additionally, co-expression analyses identified genes associated with development and host invasion in unsporulated and sporulated oocysts and immune interactions during gametocyte parasitism. CONCLUSIONS: This is the first study, to our knowledge, to use the global transcriptome profiles to decipher molecular changes across the E. stiedae life cycle, and these results not only provide important information for the molecular characterization of E. stiedae, but also offer valuable resources to study other apicomplexan parasites with veterinary and public significance.


Assuntos
Coccidiose/veterinária , Eimeria/genética , Coelhos/parasitologia , Transcriptoma , Animais , Coccidiose/parasitologia , Eimeria/crescimento & desenvolvimento , Eimeria/isolamento & purificação , Eimeria/metabolismo , Merozoítos/genética , Merozoítos/crescimento & desenvolvimento , Merozoítos/metabolismo , Oocistos/genética , Oocistos/crescimento & desenvolvimento , Oocistos/metabolismo , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Análise de Sequência de RNA
13.
Int J Parasitol ; 51(11): 977-984, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34089715

RESUMO

Interplay between conserved host specificity and occasional host switches is an important process determining the evolution of host-parasite systems. Here, we address the dynamics of host switches at the population level in rodent-associated Eimeria. Focusing mainly on two ecologically similar host groups, Murinae and Arvicolinae, we show that the Eimeria infecting those hosts form a complex system of many genetic lineages with different host specificities. The broad geographic distribution of lineages indicates that they are well-established genetic forms which retained their host specificities while spreading across large geographic areas. We also demonstrate that genetic structure is only partially reflected by morphological traits.


Assuntos
Eimeria , Animais , Arvicolinae , Especificidade de Hospedeiro , Interações Hospedeiro-Parasita , Humanos , Murinae , Filogenia
14.
Vet Parasitol ; 296: 109480, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34120030

RESUMO

Eimeria necatrix is one of the most pathogenic chicken coccidia and causes avian coccidiosis, an enteric disease of major economic importance worldwide. Eimeria parasites have complex developmental life cycles, with an exogenous phase in the environment and an endogenous phase in the chicken intestine. Oocysts excreted by chickens rapidly undergo meiosis and cell division to form eight haploid sporozoites (SZ). SZ liberated from sporocysts in the chicken intestine migrate to their preferred site of development to initiate cellular invasion. To date, almost nothing is known about the proteins that mediate parasite invasion in E. necatrix. In order to discover genes with functions involved in cellular invasion, the transcriptome profiles of E. necatrix unsporulated oocysts (UO) and SZ were analyzed using a combination of third-generation single-molecule real-time sequencing (TGS) and second-generation sequencing (SGS) followed by qRT-PCR validation. Correction of TGS long reads by SGS short reads resulted in 34,932 (UO) and 23,040 (SZ) consensus isoforms. After subsequent assembly, a total of 4949 and 4254 genes were identified from UO and SZ libraries, respectively. A total of 8376 genes were identified as differentially expressed genes (DEGs) between SZ and UO. Compared to UO, 4057 genes were upregulated and 4319 genes were downregulated in SZ. Approximately 1399 and 1758 genes were defined as stage-specific genes in SZ and UO, respectively. Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that 2978 upregulated SZ genes were clustered into 29 GO terms, and 857 upregulated SZ genes were associated with 26 KEGG pathways. We also predicted a further 50 upregulated SZ genes and 73 upregulated UO genes encoding microneme proteins, apical membrane antigens, rhoptry neck proteins, rhoptry proteins, dense granule proteins, heat shock proteins, calcium-dependent protein kinases, cyclin-dependent kinases, cGMP-dependent protein kinase, and glycosylphosphatidylinositol-anchored surface antigens. Our data reveal new features of the E. necatrix transcriptional landscape and provide resources for the development of novel vaccine candidates against E. necatrix infection.


Assuntos
Eimeria , Interações Hospedeiro-Parasita , Oocistos , Esporozoítos , Transcriptoma , Animais , Galinhas , Eimeria/genética , Interações Hospedeiro-Parasita/genética , Oocistos/genética , Análise de Sequência de RNA/veterinária , Esporozoítos/genética , Transcriptoma/genética
15.
Vet Parasitol ; 296: 109443, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34147767

RESUMO

DNA-based diagnostic assays for detecting infections with Eimeria species have been limited to providing identification and presence/absence data for samples containing oocysts. Modern technologies that generate quantitative data, such as droplet digital PCR (ddPCR) and Next Generation Sequencing (NGS), utilize a relatively short amplicon size containing sufficient species-specific variation for reliable species level identification. Targeting the cytochrome c oxidase subunit III gene in the mitochondrial genome, we established protocols using these technologies to determine the relative abundance of the number of copies/µL of Eimeria species in a sample. Samples from chickens of known and unknown Eimeria species composition were analyzed to determine the suitability of these technologies as diagnostic assays. All technologies demonstrated robust capability of identifying and quantifying the Eimeria species in samples. The new quantitative assays described herein will produce invaluable detail of Eimeria species infections for an array of situations in commercial chicken production systems, enabling further characterization of the disease profile and allowing for the development or enhancement of new intervention strategies.


Assuntos
Coccidiose , DNA , Eimeria , Sequenciamento de Nucleotídeos em Larga Escala , Doenças das Aves Domésticas , Animais , Galinhas , Coccidiose/parasitologia , Coccidiose/veterinária , DNA/análise , DNA/química , Eimeria/genética , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/parasitologia
16.
Rev Bras Parasitol Vet ; 30(2): e026920, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34076057

RESUMO

The objective of this research was to evaluate the control of coccidiosis in the litter of commercial poultry housing in the Western region of Paraná, Brazil, which adopts sanitary management to control Salmonellosis. Three litter treatments were selected, proposed by an integrating company in the region: fermentation associated with liming (FL); liming (L) and fermentation (F). Stool samples were collected from 18 poultry housing, in a completely randomized experimental design, testing different collection areas in the sheds, from the 5th day of the birds' housing, repeated weekly until the 40th day, during five successive flocks. The incidence of coccidiosis was determined by oocysts count in feces (OoPG), testing the interactions between treatment, age of birds and the number of flocks housed. Samples were selected to identify, through the Multiplex PCR, species of the genus Eimeria. circulating in the shed. The results showed that in litter where fermentation was adopted, the efficiency to control the sporulation of Eimeria spp. was significantly (p <0.05) higher than in the other treatments. The use of the Multiplex PCR technique requires improvements in the preparation of samples for the extraction of genetic material.


Assuntos
Coccidiose , Coccidiostáticos , Eimeria , Doenças das Aves Domésticas , Animais , Brasil , Galinhas , Coccidiose/tratamento farmacológico , Coccidiose/prevenção & controle , Coccidiose/veterinária , Coccidiostáticos/uso terapêutico , Habitação , Aves Domésticas , Doenças das Aves Domésticas/prevenção & controle
17.
Parasite ; 28: 48, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34076575

RESUMO

The gastrointestinal tract in poultry harbours a diverse microbial community that serves a crucial role in digestion and protection. Disruption of the gut environment due to Eimeria spp. parasite infection causes an imbalance in intestinal homeostasis, driving the increment of pathogens such as Clostridium species. Coccidiosis infection affects the composition and integrity of gut microbiota, resulting in elevated susceptibility to diseases that pose a serious threat to the overall health and productivity of chickens. Anticoccidial drugs have proven effective in curbing coccidiosis but with concerning drawbacks like drug resistance and drug residues in meat. The exploration of natural alternative strategies such as probiotics and phytochemicals is significant in controlling coccidiosis through modification and restoration of gut microbiota, without inducing drug resistance. Understanding the interaction between Eimeria parasites and gut microbiota is crucial for the control and prevention of coccidiosis, and the development of novel alternative treatments.


Assuntos
Coccidiose , Eimeria , Microbioma Gastrointestinal , Doenças das Aves Domésticas , Animais , Galinhas , Coccidiose/prevenção & controle , Coccidiose/veterinária , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/prevenção & controle
18.
Parasitol Res ; 120(7): 2689-2693, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34146126

RESUMO

Poultry coccidiosis causes considerable economical losses to the livestock industry. Eimeria parasites are responsible for this disease. On a global scale, E. acervulina and E. tenella are amongst the most common Eimeria spp. infecting broilers. E. tenella is commonly used as infection model in in vivo and in vitro studies. On the other hand, E. acervulina has barely been studied under in vitro conditions. A well established and widely used in vitro model for E. tenella infection is the Madin-Darby bovine kidney cell line (MDBK); however, little is known regarding suitability of MDBK cells as host cells for E. acervulina. We infected MDBK monolayers with two different doses, 5 × 104 and 2 × 105, of E. acervulina sporozoites and evaluated cultures at 24 and 96 h post infection (hpi). For comparison, we ran an identical infection assay using E. tenella sporozoites. To assess parasite reproduction, the number of DNA copies of E. acervulina SCAR marker and E. tenella ITS-1 gene was quantified using real-time quantitative PCR. We found that the number of E. acervulina copies increased significantly at 24 hpi in comparison to E. tenella (p < 0.05). After 96 hpi, E. acervulina gene copies were considerably reduced while E. tenella continued to multiply (p < 0.05). Our results show that MDBK monolayers could be used for in vitro research aimed to study E. acervulina sporozoite cell invasion. Nevertheless, modifications of in vitro cultivation appear necessary to allow qualitative and quantitative studies over longer periods of parasite reproduction.


Assuntos
Eimeria/fisiologia , Rim/parasitologia , Animais , Bovinos , Linhagem Celular , Galinhas/parasitologia , Coccidiose/parasitologia , Coccidiose/veterinária , Eimeria/classificação , Eimeria/genética , Eimeria tenella/genética , Eimeria tenella/fisiologia , Células Epiteliais , Rim/citologia , Doenças das Aves Domésticas/parasitologia , Reação em Cadeia da Polimerase em Tempo Real , Esporozoítos/classificação , Esporozoítos/genética , Esporozoítos/fisiologia
19.
Poult Sci ; 100(8): 101233, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34174568

RESUMO

Rotation with different active ingredients is among the most effective and recommended strategies to preserve the efficacy of anticoccidial drugs and reduce the emergence of resistance. Tools such as anticoccidial sensitivity tests (ASTs) are ideally used to make rational rotation programs and bring benefits to production. The objective of this study was to evaluate the sensitivity of E. acervulina (EA) and E. maxima (EM) from 3 different regions in Brazil, by using four ASTs. Feces samples weighing 6 to 7 kg were collected in the regions of São Paulo, Paraná, and Minas Gerais. Prevalent oocysts from feces were filtered, identified, and quantified to conduct 2 ASTs with EA and 2 with EM. The same experimental design was used in every AST (4 replicates per treatment, with 6 birds each, for a total of 240 birds). Treatment groups were a nonchallenged and nonmedicated control group (T1), a challenged and nonmedicated control group (T2), and the other groups challenged and treated with the following compounds: lasalocid (90 ppm - T3), maduramycin (6 ppm - T4), decoquinate (30 ppm - T5), nicarbazin+semduramicin (66 ppm - T6), monensin (110 ppm - T7), salinomycin (66 ppm - T8), narasin+nicarbazin (100 ppm - T9), and nicarbazin (125 ppm - T10). At the end of each AST (20 d), the percent change (delta value) between the treated group (T3 to T10) and the control group (T2) was calculated for the following variables: body weight gain, feed conversion ratio, lesion score, and an indicator of percentage of optimal anticoccidial activity (POAA) that included T2. Different sensitivity levels of EA and EM isolates could be identified. As a whole, drugs from T5 and T3 groups showed higher delta values when compared to other compounds, whereas the lowest sensitivity levels of these isolates were observed in groups T4 and T7. Despite some limiting factors, ASTs can be a good tool for strategic selection of anticoccidial drugs in order to maintain efficacy and extend the lifespan of these molecules.


Assuntos
Coccidiose , Coccidiostáticos , Eimeria , Preparações Farmacêuticas , Doenças das Aves Domésticas , Animais , Brasil , Galinhas , Coccidiose/tratamento farmacológico , Coccidiose/veterinária , Coccidiostáticos/farmacologia , Coccidiostáticos/uso terapêutico , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/epidemiologia
20.
Front Immunol ; 12: 621803, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34149685

RESUMO

Feeding practices have been found to influence gut microbiota which play a major role in immunity of poultry. In the present study, changes in cecal microbiota and humoral responses resulting in the 55 ppm bacitracin (BACI), 1% each of cranberry (CP1) and wild blueberry (BP1) pomace alone or in combination (CP+BP) feeding in broiler Cobb 500 vaccinated or not against coccidiosis were investigated. In the non-vaccinated group, no significant treatment effects were observed on performance parameters. Vaccination significantly affected bird's performance parameters particularly during the growing phase from 10 to 20 days of age. In general, the prevalence of coccidiosis and necrotic enteritis (NE) was reduced by vaccination (P < 0.05). BACI-treated birds showed low intestinal lesion scores, and both CP1 and BP1 feed supplementations reduced Eimeria acervulina and Clostridium perfringens incidences similar to BACI. Vaccination induced change in serum enzymes, minerals, and lipid levels in 21-day old birds while, levels of triglyceride (TRIG) and non-esterified fatty acids (NEFA) were higher (P < 0.05) in CP1 treated non-vaccinated group than in the control. The levels of NEFA were lower in BACI- and CP1-fed birds than in the control in non-vaccinated day 28 old birds. The highest levels of all estimated three immunoglobulins (IgY, IgM, and IgA) were found in the vaccinated birds. Metagenomics analysis of the cecal bacterial community in 21-day old birds showed the presence of Firmicutes (90%), Proteobacteria (5%), Actinobacteria (2%), and Bacteroidetes (2%). In the vaccinated group, an effect of BACI was noted on Proteobacteria (P = 0.03). Vaccination and/or dietary treatments influenced the population of Lactobacillaceae, Enterobacteriaceae, Clostridiaceae, and Streptococcaceae which were among the most abundant families. Overall, this study revealed that besides their beneficial effects on performance, alike bacitracin, berry pomaces in poultry feed have profound impacts on the chicken cecal microbiota and blood metabolites that could be influenced by vaccination against coccidiosis.


Assuntos
Infecções Bacterianas/imunologia , Doenças das Aves/imunologia , Ceco/microbiologia , Galinhas/imunologia , Coccídios/fisiologia , Coccidiose/imunologia , Eimeria/fisiologia , Microbioma Gastrointestinal/imunologia , Vacinas Protozoárias/imunologia , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Bacitracina , Mirtilos Azuis (Planta) , Imunidade Humoral , Metabolismo dos Lipídeos , Vacinação , Vaccinium macrocarpon
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