Pharmacological Effects of Grape Leaf Extract Reduce Eimeriosis-Induced Inflammation, Oxidative Status Change, and Goblet Cell Response in the Jejunum of Mice.

Coccidiosis is a parasitic infection threatening poultry products globally. Parasite resistance to drugs is one of the barriers to Eimeria control. Natural products are one of the sources of compounds that prevent parasite infections. The current study was, therefore, conducted to evaluate the effect of Vitis vinifera leaf extract on anti-inflammatory response, oxidative status, and goblet cell response against Eimeria papillate infection in mice. Methanol was used as a solvent for phytochemicals. The mice were divided into six groups: The first group was the control. The second group was uninfected and treated with 200 mg/kg of extract to test toxicity, and the third, fourth, fifth, and sixth groups of mice received 1 × 103 sporulated E. papillate oocysts. The third group received no treatment. The fourth and fifth groups were treated daily with 100 and 200 mg/kg of V. vinifera leaf extract, respectively, while the sixth group received 25 mg/kg of toltrazuril daily via gavage. On day 5 p.i., the animals were sacrificed, and jejunum samples were prepared for analyses of histological sections and oxidative stress. The phytochemical analysis using GC-MS of the extract showed the presence of 12 biologically active compounds. The most effective dose was 200 mg/kg, which significantly decreased the number of parasitic stages in the jejunal sections of the mice. The findings demonstrate that E. papillate infection in mice results in significant histopathological changes in the jejunum, including inflammation, epithelial vacuolation, villi loss, and a decrease in goblet cell density. When infected mice received treatment, the histological injury score within the infected jejunum tissue decreased by 63%, and the goblet cell quantity dramatically increased, approaching the control values. Finally, the extract ameliorated the changes in glutathione and malondialdehyde due to E. papillate infection. The extract was proven to have anti-inflammatory properties and reduce the number of oocysts. Overall, the findings show that V. vinifera leaf extract has significant anticoccidial effects in vivo.

Melatonin Administration to Pregnant Ewes for Coccidiosis Control in Their Offspring.

In livestock research, there has been a growing interest in the impact of melatonin on both health and disease conditions. The hypothesis of the present study was that melatonin treatment prenatally could support the immune competence and growth of experimentally infected lambs. This is the first study that aimed to investigate the impact of melatonin administration throughout pregnancy on immunity and oocyst excretion of pre-partum ewes and their offspring after experimental infection with Eimeria species. Thirty pregnant ewes were allocated into five equal groups, ΚΜ, ΚC, CM, CC, and NC, and gave birth to 47 lambs. Ewes of the KM and KC groups were orally challenged with a cocktail of Eimeria-sporulated oocysts (mainly consisting of Eimeria ovinoidalis), on day 120 of pregnancy, as well as all the lambs at the age of 5-9 days apart from those born from the NC group (environmental control). Fecal samples were collected from all ewes before infection and at parturition and from all lambs 14 times (S0-S13), before infection and during the following 8 weeks, for counting oocysts per gram of feces (OPG). Immunoglobulin (IgG) and cytokine (IL-1ß, IL-6, IL-10, IFN-γ) levels were determined in ewes' plasma collected before infection and at parturition, in lambs' plasma at 24 and 72 h after their birth, and in colostrum samples at parturition and 72 h later. Body weight of lambs was recorded five times from birth until the age of 60 days. Accordingly, the leucogram was evaluated in blood samples collected six times within the same period. On average, IgG concentration was higher (p < 0.05) in the blood of KM-ewes compared to KC and CC groups and in colostrum of KM-ewes compared to other groups (p < 0.001). KM-lambs had greater IgG titer and IFN-γ level than the other groups (p < 0.05). The IL-10/ IFN-γ ratio in KM-ewes was lower than the CC group (p = 0.06). Overall, the growth rate of lambs did not differ among groups (p > 0.05). Total oocysts' excretion in KM- and CM-lambs was reduced by 94.9% (p = 0.05) and 92.6% (p = 0.025), respectively, compared to KC-lambs, following the 3-week period after challenge, when E. ovinoidalis predominated in all groups. The dominant type of leucocytes was monocytes in all experimentally infected lambs, but not in NC-lambs, while overall lymphocytes were lower in KC-lambs than in NC-lambs (p < 0.05). Considering that almost all young indoor-reared lambs are exposed to coccidia species during their early life, melatonin treatment prenatally could suggest an alternative management tool in alleviating infection pressure.

The Anticoccidial Effect of Alcoholic Vitis vinifera Leaf Extracts on Eimeria papillate Oocysts Isolated in Mice In Vitro and In Vivo.

Eimeria spp. causes eimeriosis in the guts of numerous domestic mammals and poultry, and the employment of medication and the effects of certain aspects of synthetic anticoccidials in the treatment of eimeriosis have given rise to the appearance of resistant parasites that require the search for alternate remedies. Natural products, which are safe and have no negative impact on the environment, may be utilized in the therapy of an enormous range of parasitic infections. This research aimed to assess the effectiveness of VVLE on the oocyst sporulation of an E. papillate infection in the mouse jejunum. In addition, obtaining the ideal concentration will interrupt the parasite's life cycle and limit infection. In vitro: Collected unsporulated oocysts (1 × 103) of E. papillata were given six different concentrations (w/v) of Vitis vinifera leaf extract (10, 25, 50, 100, 150, and 200 mg/mL) and toltrazuril (25 mg/mL), three replicates per group, whereas the control group received 2.5% potassium dichromate solution. In vivo: The mice were separated into six groups; the first and second groups did not receive infection, whilst the third, fourth, fifth, and sixth groups were each given 1 × 103 sporulated oocysts of E. papillate in the experiment. In addition, an oral dosage of 100 and 200 mg/kg VVLE were given to the fourth and fifth groups, while the sixth group was given toltrazuril at 25 mg/kg. On the fifth day, unpopulated oocysts were collected from each mouse separately. The incubation period and treatments had considerable impacts on the rate of sporulation. The infrared spectroscopy of V. vinifera extract revealed many expected active classes of chemical compounds. Further, the infection of mice with E. papillata caused an oocyst output of nearly 2 × 104 oocysts/g of faeces. VVLE significantly decreased the oocyst output to nearly 88%. In addition, we detected an inhibitory effect on the sporulation (%) and harm (%) of E. papillata oocysts in a dosage-dependent modality compared with the control group. Furthermore, they destroyed the oocyst morphology in terms of the shape, size, and quantity of sporocysts. The results indicate that grape vines have powerful activity as anticoccidials.

Prevalence and diversity of Eimeria spp. in free-range chickens in northeastern Brazil.

In tropical regions, family farming is a form of production and work that is highly present in rural areas. Because the production system for free-range chickens has a low level of technification, it frequently presents massive infection by coccidia. The objective of this study was to determine the prevalence and diversity of Eimeria species in free-range chickens in northeastern Brazil. Fecal analyses were carried out using materials collected from 100 farms, belonging to 10 different municipalities. The sample from each farm was composed of five stool samples collected from different animals. Coproparasitological analyses were performed and, in each positive sample, photomicrographs of 20 oocysts were used for morphological identification of coccidia. The presence of Eimeria spp. was detected in 59% (59/100) of the farms analyzed. Species identification was performed through morphometric analysis of 1,180 sporulated oocysts. The following eight species of Eimeria spp. were found, in decreasing order of prevalence: Eimeria necatrix (25%), Eimeria mitis (18.3%), Eimeria mivati (17.3%), Eimeria tenella (12.4%), Eimeria brunetti (9.9%), Eimeria acervulina (9.1%), Eimeria praecox (4.8%) and Eimeria maxima (3.2%). The prevalence and diversity of Eimeria spp. on farms producing backyard chickens in the semiarid region of Brazil were high, especially the diversity of species. Changing the management, with the adoption of sanitary measures, may be effective in reducing the high prevalence of coccidia on the farms studied.

Therapeutic effects of Hirudo medicinalis extract antigens on modulation of CD4+CD25+Foxp3 T cell activity in murine eimeriosis.

Eimeriosis is a common parasitic disease in the chicken industry. The aim of this study was to assess the protective role of Hirudo extract antigens (HEA) against murine eimeriosis induced by Eimeria papillate. The oocyst output, developmental stages, goblet cells and oxidative stress, were investigated. Immunohistochemistry was used to detect anti-apoptotic Bcl2 marker and the number of both CD4+ and CD25+ cells in jejunal tissue, while ELISA was used to quantify TGF-ß, IL-10 and IL-22 in jejunal tissue homogenate. Real-time PCR was also used to detect mRNA expression of mucin 2 (MUC2), inducible nitric oxide synthase (iNOS), IL-1ß, IFN-γ, TNF-α, IL-6, and FoxP3. The most effective dose (5 µg/mice) reduced the oocyst output by 82.95 ± 1.02% (P ˂ 0.001). Similarly, the same dose reduced the jejunal developmental stages by 66.67 ± 0.49% (P ˂ 0.001). Furthermore, HEA therapy increased the number of jejunal goblet cells by 12.8 ± 1 (P ˂ 0.001) and the expression of MUC2 by 0.83 ± 0.06 (P ˂ 0.001). In contrast, TNF-α, IFN-γ, IL-6, iNOS, and IL-1ß expression as well as apoptosis were reduced. The number of CD4+ and CD25+ in the jejunal tissue was increased (14.6 ± 1.2 (P ˂ 0.001), 6.84 ± 1 (P ˂ 0.01), respectively) after HEA therapy. The molecular analysis showed an increased expression of intestinal Foxp3 (3.2 ± 0.13 (P ˂ 0.001), while IL-22 was reduced (124 ± 10 (P ˂ 0.001)) versus an increase in TGF-ß (250 ± 17 (P ˂ 0.01)) and IL-10 (236 ± 16 (P ˂ 0.001)) after HEA treatment in comparison to the non-treated infected group. With respect to the infected group, HEA reduced lipid peroxidation (LPO) (15.7 ± 1.12 (P ˂ 0.001)) and nitric oxide (NO) (13 ± 1.3 (P ˂ 0.001)) but increased reduced glutathione (GSH) (3.7 ± 0.26 (P ˂ 0.001)). In conclusion, HEA therapy protected against intestinal tissue damage by activation of CD4+CD25+Foxp3 cells which showed anti-inflammatory action. Hence, HEA can be recommended as a therapeutic treatment for eimeriosis.

Long-term efficacy of toltrazuril in naïve calves prophylactically treated and experimentally infected with Eimeria spp.

Eimeria are ubiquitous parasites and eimeriosis treatment is based on coccidiostats or coccidicides used prophylactically, metaphylactically, or therapeutically. The long-term efficacy of toltrazuril (TZR, 15 mg/kg) against experimentally infected naïve calves was investigated. Seven groups (six treated and one control) of six animals each were formed. Animals of each group received a single TZR prophylactic oral treatment on days - 42, - 35, - 28, - 21, - 14, and - 7 before the challenge with infectious sporulated oocysts of Eimeria spp. (100,000 oocysts: 59.5% E. zuernii, 38.1% E. bovis, 1.2%, E. alabamensis, and 1.2% E. ellipsoidalis). The long-term efficacy was assessed based on Eimeria spp. oocyst excretion by fecal oocyst counts. Three calves from the control group presented diarrhea with blood, which was not observed in animals belonging to the treatment groups. The TZR achieved efficacy greater than 95.0% up to 14 and 7 days. This formulation showed efficacy above 95% for 7 to 14 days, between 82 and 84% for 21 to 28 days and between 50 and 64% for 35 to 42 days.

Development of a multiplex polymerase chain reaction technique for detection and discrimination of Eimeria spp. in cattle in Indonesia.

Background and Aim: Bovine eimeriosis is a disease caused by apicomplexan parasites of the genus Eimeria. It is one of the most important and widespread bovine illnesses in the world. Some of the identified species of bovine eimeriosis have morphologically similar oocysts that are difficult to differentiate. For the identification of particular Eimeria spp., diagnostic laboratories are increasingly turning to DNA-based technology. This study aims to develop a multiplex polymerase chain reaction (mPCR) technique based on the internal transcribed spacer-1 (ITS-1) gene for the simultaneous identification of pathogenic Eimeria spp. in cattle from Sulawesi Island, Indonesia. Materials and Methods: Genomic DNA was extracted by the DNAzol reagent from the purified Eimeria oocysts. Species-specific primers targeting the ITS-1 region were used to amplify the distinct Eimeria spp. Results: Using PCR ITS-1, this study showed that 36 of 120 fecal samples (30%) were infected by Eimeria spp. The multiplex PCR assay allowed for the simultaneous identification of six major Eimeria spp. in a single-tube reaction. The proportion of mixed Eimeria spp. infections was 100% (36/36). The maximum number of Eimeria spp. was five, and the minimum number was two. Conclusion: Identification of six pathogenic Eimeria spp. in cattle was successfully carried out by nested multiplex PCR using ITS-1 gene. In the future, a procedure to detect pathogenic Eimeria spp. in one tube reaction will offer economical and save diagnostic time.

Assessment of the oxidative status and goblet cell response during eimeriosis and after treatment of mice with magnesium oxide nanoparticles.

Magnesium nanoparticles have been the focus of study over the past few years because of its functionality in the body. Assessment of the impact of magnesium oxide nanoparticles (MgNPs) on eimeriosis has yet to be conducted. The goal of this study was to see how MgNPs affected the parasite Eimeria papillata infected jejunum. To induce eimeriosis, mice were infected with sporulated oocysts. For treatment, 5 mg / Kg MgNPs was used for 5 consecutive days. The infection reduced the number of intestinal goblet cells and their associated genes MUC2 and MUC4, as well as increasing oxidative damage in the jejunum. MgNPs significantly reduced the oocyst production in the feces by about 77 %. After treatment, the number of goblet cells per villus increased from 4.17% to 7.40.6%. Moreover, the MgNPs were able to upregulate the expression of MUC2 and MUC4-mRNA. MgNPs significantly increased the activity of catalase and superoxide dismutase, as well as the extent of glutathione, by day 5 after infection with the parasite. On contrary, MgNPs decreased the level of malondialdehyde and nitric oxide. The findings suggested that MgNPs could be an effective anti-eimeriosis agent due to their anti-eimerial and anti-oxidant roles, as well as the regulatory effect on the goblet cell mucin genes in the jejunum of mice.

Eimeria spp. in naturally infected beef cattle: Dynamics of oocysts excretion, prevalence, and comparison between parasitological diagnostics.

Eimeria spp. infections cause mortality, reduced well-being, and substantial economic losses implications for cattle production worldwide. The present work followed up the excretion of Eimeria spp. oocysts in two naturally infected beef herds, from two different properties, to investigate the dynamics of oocyst excretion and the prevalence of Eimeria spp. in different animal categories and seasons of the year (rainy season - October to April; dry season - May to September). Even that, the species of Eimeria were identified and the parasitological techniques of Gordon and Whitlock modified and Mini-FLOTAC were used. In both herds, animals up to 14 months had a mean total OPG counts higher than older animals (after 15-16 months of age), and the species E. zuernii and E. bovis were more frequently identified, the first species being more frequent in animals from 1 to 2 months of age, while E. bovis prevailed from three months old. On property 1, the highest mean OPG counts (P ≤ 0.05) were obtained between October 2017 and September 2018, with the highest mean OPG counts in October 2017, when the animals were aged between 4-5 months. The prevalence of the pathogen on property 1 was 59.16 % and 43.62 % in the rainy and dry season, respectively, a higher parasitic load (P ≤ 0.05) was verified in the rainy season. On property 2, the mean OPG counts of Eimeria spp. was higher (P ≤ 0.05) in animals between 8-16 months, with the highest peak in November 2019, when they were one year old. The on-site prevalence during the rainy season on property 2 was 53.09 % and 49.79 % on dry season, and no difference (P = 0.92) in the mean OPG counts of Eimeria spp. during the seasons. There was a difference (P ≤ 0.05) in the count of oocysts in females after 18 months of age than males, which was probably due to the increase in animal density. Both tested techniques can be used for quantification of the excretion of oocysts of Eimeria spp. in cattle feces showing the same OPG mean count (r = 0.9287; p = 0.0025; R² = 0.8625). Mini-FLOTAC showed higher prevalence for Eimeria spp., however, can be an obstacle depending on the number of fecal samples that need to be processed.

Efficiency of 5% toltrazuril in the metaphylactic treatment of coccidiosis in naturally infected and extensively reared beef calves.

Eimeria spp. infection is one of the main diseases that affect young cattle in beef and dairy herds, leading to economic losses and compromising the performance and growth of animals. This study aimed to evaluate the effectiveness of different protocols of metaphylactic treatment with 5% toltrazuril (15 mg/kg) in naturally infected, extensively reared beef calves from birth to 1 year of age. We used 92 calves aged up to 2 weeks of life, initially divided into two experimental groups: treated and control. The animals were evaluated fortnightly with individual count of oocysts per gram of feces (OPG) tests until weaning; thereafter, they were again divided into four subgroups: I-treated at both birth and weaning; II-treated at birth but not treated at weaning; III-not treated at birth but treated at weaning; and IV-neither treated at birth nor at weaning (control). During the experimental period, the animals were evaluated for weight gain, excretion of oocysts, and presence of pathogenic species of Eimeria spp. In the first stage of the study, less oocysts were excreted in the treated group for up to 75 days and at 150 days of age (p < 0.05). In the final stage of the study, significant differences occur between groups I and II, I and IV, II and III, and III and IV (p = 0.3512), in the collection 20 days after treatment. Eimeria bovis was the most frequently identified species during the experimental period. Moreover, there was a positive correlation between the presence of diarrhea in the calves and the number of oocysts excreted, treated group (p = 0.0092) and untreated group (p = 0.2). There was no significant difference in weight gain of animals in any of the study periods (p > 0.05).

Protective Role of Morus nigra Leaf Extracts against Murine Infection with Eimeria papillata.

BACKGROUND: Eimeriosis is a parasitic intestinal infection that affects the poultry industry; it is responsible for economic losses on a global scale. OBJECTIVE: This study investigated the protective role of Morus nigra leaf extracts (ME) against Eimeria papillata infection in mice. METHODS: C57Bl/6 mice were divided into six groups. The first group was gavaged daily with 100 µL 0.9% NaCl. The second group was treated daily by oral gavage with 100 µL M. nigra leaf extracts (ME) (200 mg/kg), while the third, fourth, fifth, and sixth groups were orally infected with 1000 E. papillata oocysts. The last three groups were daily treated for 5 days with 200, 400, and 800 mg/kg of ME, respectively. Samples of jejunum were obtained for examining the histopathological changes as well as changes in the nitric oxide and catalase levels. RESULTS: Infrared spectroscopy of ME showed the presence of several expected active classes of phytochemical compounds. ME was able to reduce the E. papillata oocysts in mice feces by about 80% and decrease the infection-induced jejunal histopathological injuries. This was quantified using the histological injury score. In addition, ME significantly recovered the changes in nitric oxide and catalase levels. CONCLUSION: Our findings showed that the M. nigra extract has an antioxidant activity which protects against murine eimeriosis. Further studies are required to determine the exact active compounds in ME and their mode of action.

A 10-Year Surveillance of Eimeria spp. in Cattle and Buffaloes in a Mediterranean Area.

Coccidiosis due to Eimeria spp. are widespread parasitic infections in cattle and water buffaloes and may impair health, welfare, and production of these livestock species. The aims of this study were (i) to investigate the prevalence and seasonal dynamics of eimeriosis and (ii) to characterize the Eimeria species in large ruminants in a Mediterranean area, in order to plan effective control strategies. Parasitological data were obtained from a 10-year surveillance program (2010-2019) on 3,631 farms (2,089 buffalo and 1,542 cattle farms) sampled in central and southern Italy. Pooled fecal samples were analyzed using the FLOTAC technique with an analytic sensitivity of 2 oocysts per gram of feces (OPG) utilizing a saturated sodium chloride flotation solution (specific gravity = 1.200). Eimeria species identification was performed by morphometric analysis after a one week incubation of oocysts in a 2.5% potassium dichromate solution. The results showed high prevalence of Eimeria (up to 100%) in both cattle and buffaloes in the 10 years of surveillance, even if a slight reduction was reported in the last three years. The overall prevalence of eimeriosis was 91.7% (95% confidence interval, 95% CI = 90.2-93.1) in cattle farms and 81.5% (95% CI = 79.8-83.1) in water buffalo farms. The mean OPG value was 66.8 (min = 2; max = 8,065) in cattle and 55.9 (min = 2; max = 15,415) in water buffaloes, but this difference was not statistically significant (p > 0.05). In total, nine species of Eimeria were found in cattle the most prevalent being Eimeria bovis, E. ellipsoidalis, E. cylindrica, and E. zuernii, whereas in water buffaloes eight species of Eimeria were found, the most prevalent being E. ellipsoidalis, E. auburnensis, E. bovis, and E. zuernii. Mixed infections were common in both ruminant species. The seasonal pattern showed a higher prevalence of eimeriosis in cattle in spring (86.9%) whereas in buffalo farms the prevalence was higher in winter (82.3%) and summer (82.4%). In conclusion, the 10-year surveillance program indicates that eimeriosis is common in cattle and water buffaloes and therefore continuous effective control strategies are needed.

Ocorrência de espécies do gênero Eimeria Schneider, 1875, em cordeiros confinados / Occurrence of Eimeria Schneider, 1875 species in feedlot lambs

Este trabalho descreve as espécies do gênero Eimeria Schneider, 1875, que ocorreram em um confinamento de cordeiros, bem como as dinâmicas da eliminação de oocistos no ambiente, a correlação com o ganho de peso médio diário (GMD) e as variáveis climáticas, durante nove semanas. Cento e quatro cordeiros de diversas raças e cruzas, com aproximadamente 60 dias de vida, foram confinados e submetidos a pesagens e avaliações clínicas e coprológicas periódicas. Amostras de fezes com mais de 500 oocistos de Eimeria por grama de fezes (OoPG) foram separadas para esporulação e identificação das espécies. Entre os oocistos avaliados, foram identificadas as espécies: E. parva, E. crandallis, E. ovinoidalis, E. weybridgensis, E. bakuensis, E. marsica, E. ahsata, E. granulosa, E. pallida e E. faurei. Eimeria crandallis foi a mais frequente, presente em 44 das 58 amostras avaliadas, enquanto E. parva foi a mais abundante nas contagens individuais. Nenhum dos animais apresentou quadro de eimeriose, e coeficientes negativos foram encontrados nas correlações OoPG vs. GMD (-0,075) e OoPG vs. pluviosidade (-0,1164), enquanto para OoPG vs. temperatura foi encontrado coeficiente positivo (0,2914). Animais positivos para a eliminação de oocistos apresentaram infecção mista nas avaliações semanais, com até sete espécies parasitando um mesmo cordeiro.(AU)

This study describes the Eimeria Schneider, 1875 species that occurred in a lamb feedlot, as well as the dynamics of oocyst output in the environment and its correlation with daily weight gain (DWG) and climatic variables during nine weeks. One hundred and four lambs of various breeds and crossbreeds, at approximately 60 days old, were placed in a feedlot and submitted to periodic weighing and clinical and coprological evaluations. Fecal samples presenting more than 500 Eimeria spp. oocysts per gram (OPG) were separated for sporulation, and oocysts were identified at species level. Among evaluated oocysts, the following species were identified: E. parva, E. crandallis, E. ovinoidalis, E. weybridgensis, E. bakuensis, E. marsica, E. ahsata, E. granulosa, E. pallida and E. faurei. Eimeria crandallis was the most frequent one, being identified in 44 of the 58 evaluated samples, while E. parva was the more abundant in individual counts in most weeks. None of the animals presented clinical signs of eimeriosis and negative correlation coefficients were found for OPG vs DWG (-0.075) and OPG vs rainfall, as for OPG vs temperature this coefficient was positive. Animals shedding oocysts presented mixed infection, with up to seven species parasitizing the same lamb.(AU)

Mulberry extract as an ecofriendly anticoccidial agent: in vitro and in vivo application / Extrato de amoreira como agente anticoccidiano ecologicamente correto: aplicação in vitro e in vivo

Abstract Natural products are ecofriendly agents that can be used against parasitic diseases. Eimeria species cause eimeriosis in many birds and mammals and resistance to available medications used in the treatment of eimeriosis is emerging. We investigated the in vitro and in vivo activity of Morus nigra leaf extracts (MNLE) against sporulation of oocysts and infection of mice with Eimeria papillata. Phytochemical analysis of MNLE showed the presence of seven compounds and the in vitro effects of MNLE, amprolium, DettolTM, formalin, ethanol, and phenol were studied after incubation with oocysts before sporulation. Furthermore, infection of mice with E. papillata induced an oocyst output of approximately 12 × 105 oocysts/g of feces. MNLE significantly decreased oocyst output to approximately 86% and the total number of parasitic stages in the jejunum by approximately 87%. In addition, the reduction in the number of goblet cells in the jejuna of mice was increased after treatment. These findings suggest that mulberry exhibited powerful anticoccidial activity.

Resumo Os produtos naturais são agentes ecologicamente corretos que podem ser usados ​​contra doenças parasitárias. As espécies de Eimeria causam eimeriose em muitas aves e mamíferos e a resistência aos medicamentos disponíveis usados ​​no tratamento da eimeriose está emergindo. Foram investigadas as atividades in vitro e in vivo dos extratos de folhas de Morus nigra (MNLE) contra esporulação de oocistos e infecção de camundongos com Eimeria papillata. A análise fitoquímica do MNLE mostrou a presença de sete compostos e os efeitos in vitro do MNLE, amprolium, DettolTM, formalina, etanol e fenol foram estudados após incubação com oocistos antes da esporulação. Além disso, a infecção de camundongos com E. papillata induziu uma produção de oocistos de aproximadamente 12 × 105 oocistos / g de fezes. O MNLE reduziu significativamente a produção de oocistos para aproximadamente 86%, e o número total de estágios parasitários no jejuno em aproximadamente 87%. Além disso, a redução no número de células caliciformes no jejuno de camundongos aumentou após o tratamento. Esses achados sugerem que a amoreira exibia uma poderosa atividade anticoccidiana.

Salvadora persica protects mouse intestine from eimeriosis / Salvadora persica protege o intestino do rato da eimeriose

Abstract Eimeriosis is a global poultry health problem. In the current study, we investigated the role of Salvadora persica leaf extracts (SE) against murine eimeriosis induced by Eimeria papillata. The infection induced an oocyst output of 6242 ± 731 oocysts/g feces. After treatment with 300 mg⁄kg SE, the oocysts expelled in feces decreased by approximately 3-fold. In addition, the total number of E. papillata in the parasitic stage decreased in the jejunum of mice after treatment with SE. In addition, SE significantly reduced the number of apoptotic cells by approximately 2-fold in the infected jejunum. SE ameliorated the changes in glutathione, malondialdehyde, and catalase due to E. papillata infection. Finally, SE regulated the cytokine genes, interleukin (IL)-1β, IL-6, interferon-γ, and tumor necrosis factor-α, and the apoptotic genes, B-cell lymphoma-2, Bax, and Caspase-3. SE protects the jejunum from E. papillata induced injury and may have potential therapeutic value as a food additive during eimeriosis.

Resumo A eimeriose é um problema global de saúde avícola. No presente estudo, investigou-se o papel dos extratos de folhas de Salvadora persica (SE) contra a eimeriose murina induzida por Eimeria papillata. A infecção induziu uma produção de oocistos de 6242 ± 731 oocistos/g de fezes. Após o tratamento com 300 mg⁄kg SE, os oocistos eliminados nas fezes diminuíram em aproximadamente 3 vezes. Além disso, o número total de E. papillata no estágio parasitário diminuiu nos jejunos de camundongos após o tratamento com SE. Da mesma forma, o SE reduziu significativamente o número de células apoptóticas em aproximadamente 2 vezes no jejuno infectado. O estudo mostrou que o SE melhorou as alterações na glutationa, malonaldeído e catalase devido à infecção por E. papillata. Finalmente, o SE regulou os genes das citocinas, interleucina (IL) -1β, IL-6, interferon-γ e fator de necrose tumoral α, e os genes apoptóticos, linfoma-2, Bax e Caspase-3. Assim, o SE protegeu os jejunos das lesões induzidas por E. papillata e pode ter potencial valor terapêutico como aditivo alimentar durante a eimeriose.

Impairment of the phosphotransfer network and performance in broiler chickens experimentally infected by Eimeria spp.: The role of the oxidative stress.

The aim of this study was to evaluate whether infection Eimeria spp. in broiler chickens could negatively affect seric enzymes linked to adenosine triphosphate (ATP) metabolism and its relationship to oxidative stress. For this, 30 broiler chickens, 27 days-old, were divided into two groups (n = 15): the control group (C) and the group infected by Eimeria spp. (I). On days 1, 7 and 15 of the experiment, the animals were weighed, and fecal and blood samples were collected to evaluate the presence of oocysts and for serum biochemistry and enzymatic parameters, respectively. On day 15, one animal per repetition was submitted to euthanasia and intestinal fragments were collected for histopathological analyses. The body weight was lower in infected animals on day 15 of experiment, while oocyst counts were higher in infected animals on days 7 and 15 of the experiment. Serum levels of globulins were lower in infected animals on days 7 and 15 of experiment, while uric acid levels were higher in the same days, which represent changes on the immune system. Compared to the uninfected animals, on days 7 and 15, levels of serum globulins, triglycerides, creatine kinase and cholesterol were lower. Levels of adenylate kinase and reactive oxygen species (ROS) were higher on both days in infected animals, while levels of thiobarbituric acid-reactive substances (TBARS) were elevated on day 15. Lesions and immature forms of the parasite were observed in the intestines of infected birds. The phosphotransfer network elicited by an oxidative stress negatively affected the performance of broiler chickens with coccidiosis.

Eimeria species in dairy and beef cattle of different ages in Goiás state, Brazil / Espécies de Eimeria em bovinos de leite e corte de diferentes idades no Estado de Goiás, Brasil

Abstract This study aimed to identify the frequency of occurrence of Eimeria species in Holstein (taurine dairy cattle) and Nelore (zebu beef cattle) on 12 farms in the state of Goiás, Brazil. A total of 2,601 animals were evaluated, and the cattle were divided into the following age-group categories: up to three months old, four to eight months old, nine to 16 months old and cows. Fecal samples were collected directly from the rectum of all animals. None of the cattle presented any clinical signs of eimeriosis during the survey. From the results obtained, it could be concluded that higher rates of infection by Eimeria spp. were observed in Holstein cattle between the 1st and 16 th months of age and in Nelore cattle between the 1st and 8th months of age. Twelve and nine Eimeria species were diagnosed in Holstein and Nelore cattle, respectively. However, Eimeria zuernii was more prevalent in animals up to three months old, while Eimeria bovis was more common in the remaining categories (cattle aged four months and over). Future studies on the epidemiology and/or seasonality of infection by Eimeria species in cattle, from birth onwards, are needed in order to better understand the dynamics of E. zuernii and E. bovis occurrences, especially in cattle under three months of age.

Resumo O presente trabalho teve como objetivo identificar a frequência das espécies de Eimeria spp. em bovinos holandeses (bovinos de leite) e Nelore (bovinos de corte) em 12 propriedades no Estado de Goiás. Foi avaliado um total de 2.601 animais, e em cada propriedade, os bovinos foram divididos nas seguintes categorias conforme a idade: até 3 meses, 4 a 8 meses, 9 a 16 meses e vacas. Amostras de fezes foram coletadas diretamente da ampola retal dos animais. Com base nos resultados obtidos pode-se concluir que em holandeses e Nelores, o maior número de bovinos infectados por Eimeria spp. foi observado do 1º ao 16º e do 1º ao 8º meses de idade, respectivamente. Não houve nenhum caso de eimeriose clínica durante a pesquisa. Doze e nove espécies de Eimeria foram diagnosticadas em holandeses e Nelores, respectivamente. As duas espécies mais frequentes foram Eimeria bovis e Eimeria zuernii, entretanto, E. zuernii foi mais prevalente em animais até os 3 meses de idade, enquanto que E. bovis foi a mais frequente nas demais categorias de bovinos, a partir dos 4 meses de idade. Futuros estudos relacionados à epidemiologia e/ou de sazonalidade da infecção por espécies Eimeria spp., em bovinos desde o seu nascimento, são necessários, a fim de melhor entender a dinâmica de ocorrência de E. zuernii e E. bovis principalmente em bezerros com até 3 meses de idade.

Nanoselenium prevents eimeriosis-induced inflammation and regulates mucin gene expression in mice jejunum.

BACKGROUND: Although elemental selenium has been found to be effective against Eimeria, no study has yet investigated the effects of selenium nanoparticles (SeNPs) on the Eimeria parasite. The aim of this study, therefore, was to evaluate the ameliorative effect of SeNPs compared with elemental selenium on mice jejunum infected with sporulated oocysts of Eimeria papillata. METHODS: The mice were divided into 4 groups, with the first being the non-infected, control group, and the second, third, and fourth groups being orally inoculated with 1,000 sporulated oocysts of E. papillata. The third and fourth groups also received, respectively, an oral dose of 0.1 mg/kg sodium selenite and 0.5 mg/kg SeNPs daily for 5 consecutive days. RESULTS: The infection induced severe histopathological jejunal damage, reflected in the form of destroyed jejunal mucosa, increased jejunal oxidative damage, a reduction in the number of jejunal goblet cells, and increased production of pro-inflammatory cytokines, quantified by real-time polymerase chain reaction. Treatment of mice with SeNPs significantly decreased the oocyst output in the feces by ~80%. Furthermore, the number of parasitic stages counted in stained jejunal paraffin sections was significantly decreased after the mice were treated with SeNPs. In addition, the number of goblet cells increased from 42.6±7.3 to 95.3±8.5 cells/10 villus-crypt units after treatment. By day 5 post-infection with E. papillata, SeNPs could be seen to have significantly increased the activity of glutathione peroxidase from 263±10 to 402.4±9 mU/mL. Finally, SeNPs were able to regulate the gene expression of mucin 2, interleukin 1ß, interleukin 6, interferon-γ, and tumor necrosis factor α in the jejunum of mice infected with E. papillata. CONCLUSION: The results collectively showed that SeNPs are more effective than sodium selenite with regard to their anti-coccidial, anti-oxidant, and anti-inflammatory role against eimeriosis induced in the jejunum of mice.

Comparison between McMaster and Mini-FLOTAC methods for the enumeration of Eimeria maxima oocysts in poultry excreta.

Monitoring Eimeria shedding has become more important due to the recent restrictions to the use of antibiotics within the poultry industry. Therefore, there is a need for the implementation of more precise and accurate quantitative diagnostic techniques. The objective of this study was to compare the precision and accuracy between the Mini-FLOTAC and the McMaster techniques for quantitative diagnosis of Eimeria maxima oocyst in poultry. Twelve pools of excreta samples of broiler chickens experimentally infected with E. maxima were analyzed for the comparison between Mini-FLOTAC and McMaster technique using, the detection limits (dl) of 23 and 25, respectively. Additionally, six excreta samples were used to compare the precision of different dl (5, 10, 23, and 46) using the Mini-FLOTAC technique. For precision comparisons, five technical replicates of each sample (five replicate slides on one excreta slurry) were read for calculating the mean oocyst per gram of excreta (OPG) count, standard deviation (SD), coefficient of variation (CV), and precision of both aforementioned comparisons. To compare accuracy between the methods (McMaster, and Mini-FLOTAC dl 5 and 23), excreta from uninfected chickens was spiked with 100, 500, 1,000, 5,000, or 10,000 OPG; additional samples remained unspiked (negative control). For each spiking level, three samples were read in triplicate, totaling nine reads per spiking level per technique. Data were transformed using log10 to obtain normality and homogeneity of variances. A significant correlation (R = 0.74; p = 0.006) was observed between the mean OPG of the McMaster dl 25 and the Mini-FLOTAC dl 23. Mean OPG, CV, SD, and precision were not statistically different between the McMaster dl 25 and Mini-FLOTAC dl 23. Despite the absence of statistical difference (p > 0.05), Mini-FLOTAC dl 5 showed a numerically lower SD and CV than Mini-FLOTAC dl 23. The Pearson correlation coefficient revealed significant and positive correlation among the four dl (p ≤ 0.05). In the accuracy study, it was observed that the Mini-FLOTAC dl 5 and 23 were more accurate than the McMaster for 100 OPG, and the Mini-FLOTAC dl 23 had the highest accuracy for 500 OPG. The McMaster and Mini-FLOTAC dl 23 techniques were more accurate than the Mini-FLOTAC dl 5 for 5,000 OPG, and both dl of the Mini-FLOTAC were less accurate for 10,000 OPG counts than the McMaster technique. However, the overall accuracy of the Mini-FLOTAC dl 23 was higher than the McMaster and Mini-FLOTAC dl 5 techniques.

Surtos consecutivos ocasionados por Eimeria zuernii em bezerros de corte de uma propriedade do estado de São Paulo / Consecutive outbreaks caused by Eimeria zuernii in calves on a rural property located in the municipality of São João da Boa Vista, State of São Paulo, Brazil

O presente trabalho tem como objetivo relatar surtos consecutivos (Agosto de 2015 e 2016) ocasionado por Eimeria zuernii em bezerros de corte, Canchim e Nelore, com aproximadamente 45 dias de vida, em uma propriedade rural localizada no município de São João da Boa Vista, estado de São Paulo. O primeiro surto foi diagnosticado em um lote composto por 78 bezerros, entre 35 e 55 dias de idade. Dos 78 bezerros, 32 eram Nelore e os outros 46 Canchim. Já o segundo, ocorreu em um novo lote de 66 bezerros (sendo 36 Nelore e 47 Canchim), com idade entre 30 e 45 dias. Alguns animais apresentaram diarreia sanguinolenta e desidratação. Colheitas de fezes de todos os animais (vacas e bezerros) foram realizadas para realização de exame coproparasitológico e quantificação de ovos e oocistos por grama (OPG e OoPG) de fezes de parasitos. Com base nos resultados encontrados neste estudo, é possível afirmar que os surtos consecutivos em bezerros de corte entre 30 e 55 dias de idade, na época seca do ano (agosto), foram desencadeados pelo elevado grau de infecção destes animais por E. zuernii. Analisando os dois anos em que os surtos ocorreram, a presença de sinais clínicos (diarreia sanguinolenta) e a mortalidade de bezerros desencadeada pela eimeriose/desidratação, aconteceram em 8,3% 1,4% do rebanho, respectivamente. Estes resultados destacam a importância em se realizar pesquisas de campo, com o objetivo de se melhor entender a dinâmica de parasitismo de E. zuernii e E. bovis em bezerros com idade inferior a três meses, em função dos motivos já discutidos neste estudo. Além disso, é necessário que se investigue as possíveis fontes de infecção, como a água ingerida pelos animais, para que condutas práticas possam ser adotadas/recomendadas aos proprietários, diminuindo desta maneira os prejuízos ocasionados por surtos de Eimeria spp em rebanhos bovinos.(AU)

We report consecutive outbreaks (August 2015 and 2016) of disease caused by Eimeria zuernii in Canchim and Nelore calves about 45 days of age on a rural property in the municipality of São João da Boa Vista, state of São Paulo, Brazil. The first outbreak infected 78 calves (32 Nelore and 46 Canchim) between 35 and 55 days of age. The second outbreak infected 66 calves (36 Nelore and 47 Canchim) between 30 and 45 days of age. Some calves had bloody diarrhea and were dehydrated. Feces were collected from all animals (cows and calves) for coproparasitological examination and quantification of parasite eggs and oocysts per gram. The consecutive outbreaks in the 30 to 55 day-old calves occurred during the dry season (August). These outbreaks were caused by a high degree of infection by E. zuernii. Clinical signs (bloody diarrhea) and mortality of the calves triggered by eimeriosis/dehydration occurred in 8.3 and 1.4% of the herd in 2015 and 2016, respectively. These results highlight the importance of conducting field studies to better understand the parasitic dynamics of E. zuernii in calves younger than three months. Identifying the sources of infection, such as the water ingested by the animals, is also necessary, so that practical husbanddry can be recommended and adopted by the producers, to reduce the damage caused by outbreaks of Eimeria spp. in cattle herds.(AU)