Therapeutic efficacy of Citrus aurantifolia (lime) juice in experimental Eimeria tenella-infected broiler chickens.

The purpose of the present experiment was to explore the therapeutic effect of the crude fruit juice of Citrus aurantifolia (CAJ) on Eimeria tenella disorder in chickens. One hundred twenty 3-week-old Ross 308 broilers of equal sexes were assigned to six experimental groups of 20 birds each. Groups A, B, C, D, and E were experimentally infected with 20,000 sporulated Eimeria tenella oocysts. Broilers in groups A, B, and C were infected and allocated to three treatment-graded doses of C. aurantifolia fruit juice (20, 10, and 5 mL/kg body weight, respectively) which were administered orally for 7 consecutive days of the trial. Group D was infected and treated with a reference drug, Amprolium 1.5 g/L of drinking water (positive control), group E served as infected-untreated control, and group F was uninfected and non-treated (negative control). Oocysts per gram of feces were counted using the McMaster counting device, weight gain was calculated, and blood samples from each experimental group were collected on days 0, 7, 14, 21, and 28 post-infection for hematological evaluation. Results revealed that medication of broilers with C. aurantifolia fruit juice dose-dependently increased body weight, improved cecal lesion, decreased the E. tenella oocyst production rate, and significantly (P<0.05) increased the PCV of the infected broilers. C. aurantifolia provided valuable therapeutic effects against E. tenella-induced coccidiosis in broiler chickens. The plant fruit juice should be further validated to explore the vital compounds responsible for the anticoccidial efficacy.

Dietary organic zinc and probiotic alleviate induced Eimeria tenella infection in Japanese quails model of coccidiosis.

The current study was conducted to evaluate the impact of organic zinc (OZn) and probiotic on growth performance, oocysts number, and histological features of cecum of quails following Eimeria tenella challenge. A total of 480 Japanese quails were distributed into six treatments as follows: untreated uninfected; untreated infected; E. tenella challenge + amprolium; E. tenella challenge + OZn; E. tenella challenge + probiotic; and E. tenella challenge + OZn + probiotic. Except untreated uninfected, all groups were orally gavaged at day 8 with 5 × 104 E. tenella sporulated oocysts. Supplementation of OZn + probiotic improved (P = 0.001) growth performance compared to the untreated infected group. Lesion score of intestine and mortality was lower (P < 0.01) in quails supplemented with OZn + probiotic. Moreover, oocysts per gram (OPG) and histological dimensions of cecum in challenged birds were alleviated in OZn + probiotic. The histological findings of quails supplemented with OZn + probiotic showed normal intestinal villi with gentle sloughed epithelium. We concluded that OZn + probiotic may be safely included in the diet of Japanese quails to control coccidiosis.

Differential expression of microRNAs in the caecal content and faeces of broiler chickens experimentally infected with Eimeria.

Coccidiosis caused by Eimeria spp. incurs significant morbidity and mortality in chickens, and is thus of great economic importance. Post-mortem intestinal lesion scoring remains one of the most common means of diagnosis; therefore alternative, non-invasive methods of diagnosis and monitoring would be highly desirable. Micro-RNAs (miRNAs) have been shown to be stable in faeces of human and animal species with expression altered in gastrointestinal disease. We hypothesized that miRNA is stable in caecal content of chickens, and that differential miRNA expression patterns would be seen in Eimeria-infected versus uninfected individuals. Initially, RNA was extracted from Eimeria tenella-infected (n = 3; 7 days post infection) and uninfected (n = 3) chicken caecal content to demonstrate miRNA stability. Subsequently, next-generation miRNA sequencing was performed on caecal content from E. tenella-infected chickens with high (lesion score (LS) 3-4; n = 3) or low (LS1; n = 3) levels of pathology, and uninfected controls (n = 3). Comparative analysis identified 19 miRNAs that exhibited significantly altered expression in the caecal content of E. tenella, infected chickens versus uninfected chickens (t-test, False Discovery Rate (FDR) < 0.05). Eight of these miRNAs showed significant up-regulation in infection (fold change of 9.8-105, FDR <0.05). Quantitative PCR was performed using separate biological replicates to confirm differential regulation in eight of these miRNA candidates in caecal and faecal content. This work has identified a panel of miRNA candidates which may be appropriate for use as non-invasive faecal markers of active caecal coccidiosis without the need for culling. RESEARCH HIGHLIGHTSE. tenella induced differential miRNA expression in caecal content and faeces.

First report of Caryospora sp. infection in free-living green turtles (Chelonia mydas) in Northeastern Brazil.

Infection by Caryospora cheloniae has been reported to be responsible for green turtle strandings with high morbidity and mortality rates worldwide. Although studies have already shown the pathogenesis of these infections, many aspects of this protozoan are still poorly understood, including their life cycle and infection dynamics in free-living sea turtle populations. Due to the lack of information about the infection by this protozoan in sea turtles in Northeastern Brazil, our study aims to describe Caryospora sp. infection and its pathological findings in free-living Chelonia mydas found on the north coast of the Bahia state. Between 2018 and 2019, 64 specimens of green turtles were necropsied in partnership with Fundação Projeto Tamar; among these, 10 (1.56%) had oocysts morphologically compatible with Caryospora cheloniae in the evaluation of fecal samples and histopathological examination of intestinal samples. The infected animals were juvenile green turtles that were found stranded on the beaches of the north coast of Bahia. The pathological findings were restricted to the lower gastrointestinal tract, with different presentations and intensities. About 70% of the animals with coccidial infection exhibited erosive and ulcerative fibrinous enteritis. This is the first report of coccidiosis in green turtles on the north coast of Bahia.

Pathology caused by three species of Eimeria that infect the turkey with a description of a scoring system for intestinal lesions.

Three-week-old turkey poults were infected with pure lines of three species of Eimeria (E. adenoeides, E. gallopavonis, and E. meleagrimitis) recently isolated from commercial turkey farms. The lines had been propagated from a single oocyst and identified by species-specific PCR amplification of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Five to six days after infection their intestines were removed and examined for the presence of intestinal lesions. A description and review of the pathology caused by these parasites is provided, and a scoring system developed by which the severity of the lesions can be evaluated. The system is similar to that described by Johnson, J. and Reid, W. M. [1970. Anticoccidial drugs: lesion scoring techniques in battery and floor-pen experiments with chickens. Experimental Parasitology, 28, 30-36] for chickens in which a score of zero to four is assigned to lesions of increasing severity. The intestinal lesions observed here, and their assigned scores, are supported by representative illustrations. It is hoped that they may prove a useful tool for evaluating the pathology caused by E. adenoeides, E. gallopavonis, and E. meleagrimitis in the turkey.RESEARCH HIGHLIGHTSA scoring system has been developed for intestinal lesions caused by three species of Eimeria that infect the turkey.The lesions attributable to these species are illustrated.

Developing an experimental necrotic enteritis model in turkeys - the impact of Clostridium perfringens, Eimeria meleagrimitis and host age on frequency of severe intestinal lesions.

BACKGROUND: Necrotic enteritis is a significant problem to the poultry industry globally and, in Norway up to 30% of Norwegian turkey grow-outs can be affected. However, despite an awareness that differences exist between necrotic enteritis in chickens and turkeys, little information exists concerning the pathogenesis, immunity, microbiota or experimental reproduction of necrotic enteritis in turkeys. In particular, it is important to determine the appearance of the gross lesions, the age dependency of the disease and the role of netB toxin of Clostridium perfringens. To this end, we report our findings in developing an in vivo experimental model of necrotic enteritis in turkeys. RESULTS: A four tier (0-3) scoring system with clearly defined degrees of severity of macroscopic intestinal lesions was developed, based on 2312 photographic images of opened intestines from 810 B.U.T. 10 or B.U.T. Premium turkeys examined in nine experiments. Loss of macroscopically recognizable villi in the anterior small intestine was established as the defining lesion qualifying for a score 3 (severe intestinal lesions). The developed scoring system was used to identify important factors in promoting high frequencies of turkeys with severe lesions: a combined Eimeria meleagrimitis and Clostridium perfringens challenge, challenge at five rather than 3 weeks of age, the use of an Eimeria meleagrimitis dose level of at least 5000 oocysts per bird and finally, examination of the intestines of 5-week-old turkeys at 125 to 145 h after Eimeria meleagrimitis inoculation. Numbers of oocysts excreted were not influenced by Clostridium perfringens inoculation or turkey age. Among three different lesion score outcomes tested, frequency of severe lesions proved superior in discriminating between impact of four combinations of Clostridium perfringens inoculation and turkey age at challenge. CONCLUSIONS: This study provides details for the successful establishment of an in vivo model of necrotic enteritis in turkeys.

Establishment of a model of Neospora caninum infection in pregnant mice.

The aim of this study was to establish an animal model of Neospora caninum infection in pregnant BALB/c mice infected with different doses of N. caninum tachyzoites. After infection, the female BALB/c mice were housed with male BALB/c mice. The aim of this study was to observe clinical signs and pathological changes, detect Nc5 gene expression in the main organs, and measure the wet weight and coefficient of the placenta of the pregnant mice. In addition, the level of cytokines and placental hormones in the serum was measured in pregnant mice. Our results showed that the optimal dose of the mice in the infected model was 105 tachyzoites. The infected pregnant mice presented with various clinical signs, including depression, ataxia, and variable mortality. Pathological observations of the brain, liver, and spleen in the mice exhibited hyperemia, bleeding, and swelling. Moreover, N. caninum tissue cysts or tachyzoites were observed in the brain, liver, and spleen tissues by hematoxylin-eosin (HE). The Nc5 gene was detected in the brain, liver, spleen, and placental tissues of the mice. With the increase in infection days, the weight of the placenta in the model mice increased, and the placenta ratio decreased gradually. Compared with the control group, the placenta weight and placental ratio were significantly different (P < 0.05). Furthermore, the levels of the placental hormones, corticotropin-releasing hormone (CRH), chorionic gonadotropin (CG), prolactin (PRL), and estriol (E3), and cytokines IFN-γ, IL-4, and TGF-ß were differentially expressed between the model and the control group (P < 0.05 or P < 0.01), which indicated that infection with N. caninum caused an imbalance in the regulatory function of the placental hormones and cytokines in pregnant mice. A pregnant mouse model of N. caninum infection was successfully established in this study, providing a foundation for the study of the pathogenic mechanisms of N. caninum.

Pathological findings in genital organs of bulls naturally infected with Besnoitia besnoiti.

Bulls chronically affected by bovine besnoitiosis can suffer from sterility. There is limited information about the distribution of Besnoitia cysts and their associated lesions within the male genital organs. This work describes the gross and histological abnormalities in the genital organs of 6 bulls chronically infected with Besnoitia besnoiti, including both clinically (n = 4) and subclinically (n = 2) affected cases. Parasitic cysts were observed in the genital organs of all the clinically affected bulls. The tissue cysts were most commonly found within the pampiniform plexus (4/4), where they were often seen within venous vascular walls and associated with vasculitis, followed by epididymis (3/4), tunica albuginea (2/4), and penis (1/4). In decreasing order of their frequency, observed abnormalities included seminiferous tubule degeneration, testicular fibrosis, testicular necrosis, lack of/or diminished numbers of spermatozoa, testicular atrophy, and Leydig cell hyperplasia. Only one of the subclinically infected bulls had few Besnoitia cysts within the pampinoform plexus, which was associated to small areas of necrosis and mineralization in the ipsilateral testicle. Results indicate that Besnoitia cysts and genital abnormalities are frequent in bulls chronically affected by bovine besnoitiosis, while they are mild and scarce in subclinically affected ones. Moreover, present data show that Besnotia-associated testicular lesions can occur without the presence of cysts within the testicular parenchyma. B. besnoiti cysts seem to have a tropism for the vascular structures of the spermatic chord, which may cause testicular abnormalities via vascular damage, reduced blood flow, and/or impaired thermoregulation and subsequently lead to the observed testicular lesions.

Antioxidant and anti-apoptotic effects of selenium nanoparticles against murine eimeriosis.

Eimeriosis is caused by a protozoan parasite of the genus Eimeria and infection affecting most domestic animal species. The aim of this research was to comprehend the impact of selenium nanoparticles (SeNPs) on eimeriosis induced by Eimeria papillata in mouse jejunum, and how they work as antioxidants and anti-apoptotic agents against eimeriosis. The numbers of meronts, gamonts, and developing oocysts of E. papillata reduced after the infected mice were treated with the SeNPs. The levels of malondialdehyde (MDA), nitric oxide (NO), and other oxidative stress-related molecules, such as glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD), were assayed. E. papillata was able to change the redox status of the jejunal cells; this was confirmed by the elevation of the MDA and NO levels, and the decrease of the GSH levels and the activities of the antioxidant enzymes CAT and SOD. SeNP treatment significantly reversed this disturbance of the redox status. The expression levels of the apoptotic markers Bax and caspase-3 in the jejunal samples were evaluated using qRT-PCR. The SeNPs decreased the Bax and caspase-3 expression after being administered to the E. papillata-infected mice. Collectively, the SeNPs demonstrated antioxidant and anti-apoptotic activities against murine eimeriosis.

Effect of diet supplemented with coconut essential oil on performance and villus histomorphology in broiler exposed to avian coccidiosis.

The current research study was designed to determine the inclusion of 2% dietary essential coconut oil with and without coccidiosis challenge on performance, carcass characteristics, and intestinal histomorphology in broilers. A total of 560 broiler chicks were divided into 4 groups and then subdivided into 5 replicates. Coconut oil was used at 2% in feed, whereas coccidiosis challenged was introduced using 30,000 oocysts. The other four groups were designated as G1 (without coconut oil and without oocysts), G2 (without coconut oil with oocysts), G3 (with coconut oil without oocysts), and G4 (with coconut oil and with oocysts). The results revealed that the overall feed consumption was significantly (P < 0.01) increased in G1 and G2 than G3 and G4 groups. Overall weight gain was significantly (P < 0.01) higher in G3 compared with all other groups. Significantly (P < 0.01) better feed conversion ratio was recorded at the finisher phase in G3 and G4 groups in comparison with G1 and G2. The villus length, width, and surface area were higher (P < 0.01) in G3 compared with G2. Based on the findings of the present study, it was concluded that the use of 2% coconut oil in broiler feed improved growth performance and villus histology during coccidial challenge.

The pathogenicity of two Neospora caninum goat strains in a BALB/c mouse model.

Goats are frequently described as an intermediate host for the protozoan Neospora caninum, manifesting the disease mainly by recurrent abortions with placentitis and encephalitis in fetuses. Several reports of natural and experimental infections in cattle and mice show differences in the immune response, and the outcome of the infection can be variable depending on the species affected and by the behavior of the infective strain. This study describes for the first time two Neospora caninum strains isolated from naturally infected goats from the state of Minas Gerais, Brazil. One placenta and one brain from different goats were processed for a first bioassay in gerbils (Meriones unguiculatus). Subsequently, a second bioassay was performed by inoculating the processed brain samples from gerbils into Interferon gamma (IFN-γ) knockout mice (KO mice). Tachyzoites collected from the peritoneal fluid of the KO mice were inoculated into VERO cell monolayers, where they presented a very slow growth rate. The tachyzoites were also inoculated into BALB/c mice with a dose of 106 tachyzoites per animal. After a 5-week follow up, the animals infected with both of the strains developed a strong polarized Th1 response with increased serum and spleen gene expression levels of pro-inflammatory cytokines (mainly IFN-γ and TNF-α) in the first week. Tissue lesions were mild in the animals infected with both strains. Despite the strong immune response preventing an infection in the visceral organs, the parasite was able to reach the brain, causing progressive brain lesions from the second to fifth week post infection. The NC-goat1-infected mice presented with severe meningoencephalitis, but the NC-goat2-infected animals had considerable histological brain lesions only at week 5. Immunohistochemical analysis of the mouse brains revealed a different pattern of inflammatory cells compared to the naturally infected goats. A severe inflammatory infiltrate of CD3+ T lymphocytes was found in the NC-goat1-infected mice. A more discrete infiltrate of CD3+ T cells was found in the NC-goat2-infected animals. Additionally, IBA1 IHC revealed an intense microglial reaction and monocyte perivascular cuffs in the NC-goat1-infected animals and lower microglia/monocyte infiltrates in the NC-goat2-infected mice. This work contributes knowledge on the pathogenicity of new Neospora caninum strains in mice, comparable with other well-established mouse models of the disease, and demonstrates the importance of studying goats as an intermediate host of this parasite.

Seasonal intestinal coccidiosis in wild bluegill Lepomis macrochirus is associated with a spring bacterial epizootic.

Wild bluegill, Lepomis macrochirus Rafinesque, succumb to seasonal mortality in the early spring during cool water temperatures, shown previously to be related to bacteraemia caused by a psychrotrophic bacterium, Pseudomonas mandelii. In the study herein, intestinal coccidiosis in wild bluegill had seasonal prevalence causing heavy intestinal infections and sloughing of intestinal epithelium occurring in late winter/early spring. Infections were predominantly related to two different species, Goussia washuti n. sp., an epicellular coccidium, and a coccidium closely resembling Goussia desseri Molnár 1996, previously only described in percid fish in Europe. In 2019, co-infections of bacteraemia and intestinal coccidiosis occurred in bluegills. Evaluating coccidium infection intensity by fresh parasite examination and histology, an association was observed in which fish with moderate-to-heavy intestinal coccidiosis were 8-12 times more likely to have bacteraemia compared to fish with no or light coccidiosis. The association of these co-infections suggests that intestinal coccidiosis could contribute to seasonal bacterial epizootics of wild bluegill.

Unique regulation of Na-glutamine cotransporter SN2/SNAT5 in rabbit intestinal crypt cells during chronic enteritis.

The only Na-nutrient cotransporter described in mammalian small intestinal crypt cells is SN2/SNAT5, which facilitates glutamine uptake. In a rabbit model of chronic intestinal inflammation, SN2 stimulation is secondary to an increase in affinity of the cotransporter for glutamine. However, the immune regulation of SN2 in the crypt cells during chronic intestinal inflammation is unknown. We sought to determine the mechanism of regulation of Na-nutrient cotransporter SN2 by arachidonic acid metabolites in crypt cells. The small intestines of New Zealand white male rabbits were inflamed via inoculation with Eimeria magna oocytes. After 2-week incubation, control and inflamed rabbits were subjected to intramuscular injections of arachidonyl trifluoromethyl ketone (ATK), piroxicam and MK886 for 48 hrs. After injections, the rabbits were euthanized and crypt cells from small intestines were harvested and used. RESULTS: Treatment of rabbits with ATK prevented the release of AA and reversed stimulation of SN2. Inhibition of cyclooxygenase (COX) with piroxicam did not affect stimulation of SN2. However, inhibition of lipoxygenase (LOX) with MK886, thus reducing leukotriene formation during chronic enteritis, reversed the stimulation of SN2. Kinetic studies showed that the mechanism of restoration of SN2 by ATK or MK886 was secondary to the restoration of the affinity of the cotransporter for glutamine. For all treatment conditions, Western blot analysis revealed no change in SN2 protein levels. COX inhibition proved ineffective at reversing the stimulation of SN2. Thus, this study provides evidence that SN2 stimulation in crypt cells is mediated by the leukotriene pathway during chronic intestinal inflammation.

Comparative analysis of egg adapted vaccines and salinomycin against coccidiosis in chicks.

Avian coccidiosis is an intestinal protozoan parasitic disease of genus Eimeria. Wide use of anticoccidial drugs has resulted in development of drug resistant strains. Current study is based upon the development of egg adapted vaccines; gametocytes, formalin inactivated and formalin inactivated sonicated gametocytes against coccidiosis. Day old chicks (n = 90) were divided into six groups (A-F) with triplicate. On 5th day of age, three groups were orally given developed vaccines (0.2 ml) while on 21st day groups (A-E) were challenged orally with mixed Eimeria spp (60-70,000 oocysts/chick). The comparative effect of vaccines and drug salinomycin was evaluated on basis of different parameters. Blood, liver and caecal tissues were collected on 5th, 7th and 15th day post infection for indirect hemagglutination test, biochemical analysis and histopathology. Significantly higher body weight gain, feed consumption, mild bloody diarrhea with lowest oocyst count and survival rate of 100% was recorded for gametocytes vaccinated group (P < 0.05). Indirect hemagglutination test showed maximum antibodies titer (IgG) in gametocytes vaccinated group. Present investigation revealed that gametocyte vaccines was significantly effective in control of coccidiosis by inducing strong protection in immune chicks contained high level of antibodies that resisted heavy dose of challenge as compared to anticoccidial drug salinomycin.

Influence of dose and route of administration on the outcome of infection with the virulent Neospora caninum isolate Nc-Spain7 in pregnant sheep at mid-gestation.

Experimental infections in pregnant sheep have been focused on studying the effect of the time of challenge on the outcome of N. caninum infection, whereas the impact of the dose and route of challenge has not been studied in depth. Therefore, clinical outcome, immune responses, parasite detection and burden, and lesion severity in placental tissues and foetal brains were investigated in 90-day-pregnant sheep inoculated intravenously with 105 (G1), 104 (G2), 103 (G3), or 102 (G4) tachyzoites or subcutaneously with 104 (G5) tachyzoites of the virulent Nc-Spain7 isolate and an uninfected group (G6). Comparing challenge doses, G1 was the only group that had 100% abortion. Likewise, IFNγ levels in G1 increased earlier than those in other intravenously infected groups, and IgG levels on day 21 post-infection (pi) were higher in G1 than those in other intravenously infected groups. Concerning vertical transmission, G1 shows a higher parasite burden in the foetal brain than did G2 and G3. Comparing routes of administration, no differences in foetal survival rate or parasite load in the foetal brain were found. Although G2 had higher IFNγ levels than G5 on day 10 pi, no differences were found in humoral immune responses. Because the outcome after intravenous infection with 105 tachyzoites was similar to that observed after intravenous infection with 106 tachyzoites used in a previous work (100% abortion and vertical transmission), we conclude that it may be reasonable to use 105 tachyzoites administered by the intravenous route in further experiments when assessing drugs or vaccine candidates.

Gametogony of Eimeria macusaniensis Guerrero, Hernandez, Bazalar and Alva, 1971 in llama (Lama glama).

Camelids (llama, alpaca, vicunãs, guanacos) are important for the economy of South America and Eimeria infections are an important cause of mortality in camelids. Of the six species of Eimeria in camelids, Eimeria macusaniensis, considered the most pathogenic, is distinctive; its oocysts are the largest among all Eimeria species in animals, its prepatent period is more than 1 month, and its oocysts have been found in mummies from prehistoric times. Although, E. macusaniensis gametogonic stages are found associated with enteritis in naturally infected camelids, the schizogonic stages are unknown and clinical disease has been reported in some camelids with no oocysts in feces. Described herein are morphological details of gametogonic development and oocyst formation of E. macusaniensis in a naturally infected llama (Lama lama), solely infected with this parasite. Microgamonts, macrogamonts and oocysts were located in large (up to 300 µm diameter) parasitophorous vacuoles of enterocytes in the ileum. Schizonts were not found. Review of previous reports suggests that multinucleated microgamonts have been mistaken for schizonts. Gametogonic development described in the present study can serve as a guide for differential diagnosis of Eimeria species in the histological sections of intestines.

Epidemiological Survey on Eimeria spp. Associated with Diarrhea in Pre-weaned Native Korean Calves.

Bovine coccidiosis is one of the most important parasitic diseases affecting calf productivity. Here, we investi- gated the prevalence of Eimeria spp. in pre-weaned native Korean calves and determined the correlation between diar- rhea and Eimeria spp. Fecal samples were collected from individual calves (288 normal and 191 diarrheic) in 6 different farms. Of the 479 samples, Eimeria oocysts were detected in 124 calves (25.9%). Five Eimeria spp. were identified; E. zuernii (18.8%) was the most prevalent, followed by E. auburnensis (12.5%), E. bovis (7.5%), E. subspherica (5.8%), and E. bukidnonensis (1.0%). A significant correlation was observed between diarrhea and mixed infection with more than 2 Eimeria spp. (odds ratio [OR]=2.21; 95% confidence interval [CI]: 1.09-4.49; P=0.03) compared to single infection (OR = 1.29; 95% CI: 0.77-2.15; P = 0.33). Of the 5 Eimeria spp. identified, E. subspherica (95% CI: 1.24-5.61; P = 0.01) and E. bukidnonensis (95% CI: 825.08-1,134.25; P = 0.00) strongly increased the risk of diarrhea by 2.64-fold and 967.39-fold, respectively, compared to other species. Moreover, mixed infection with E. auburnensis and E. bukidnonensis was significantly associated with diarrhea (OR = 2,388.48; 95% CI: 1,009.71-5,650.00; P < 0.00) in pre-weaned native Korean calves. This is the first report to demonstrate the importance of E. bukidnonensis associated with diarrhea in pre-weaned native Korean calves. Further epidemiological studies should investigate the prevalence of E. bukidnonensis and the association between E. bukidnonensis and diarrhea.

Profiling of jejunum inflammatory gene expression during murine eimeriosis.

To understand the host-parasite relationship during coccidiosis it is necessary to identify the transcriptional profile of the local host. In this study, gene profiling in the mouse jejunum due to infection with Eimeria papillata was investigated using Agilent microarray technology. On day 5 post-infection, the characterization of infected and non-infected mice jejunum transcriptional response was compared. There was an increase in the level of tumour necrosis factor-α, nitrite/nitrate and nitric oxide synthase activity was observed following infection. Also, the activity of glutathione peroxidase was reduced from 86.5 to 38.2 mU/g. In addition, E. papillata infection was associated with an increase in the activities of both the mice alkaline phosphatase and lactate dehydrogenase. Moreover, experimental E. papillata infection in mice induced a significant elevation in protein carbonyl content, by about 70%. Agilent genome microarray detected 11 genes whose expression was up-regulated by more than 10-fold, and 30 genes whose expression was down-regulated by a similar amount five days after infection with E. papillata. The expression profiles of the Fas apoptotic inhibitory molecule 3(FAIM3), chemokine (C-X-C motif) receptor 5 (Cxcr5), succinate receptor 1 (SUCNR1), hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 3 (Hsd3b3) and cytochrome P450, family 2, subfamily b, polypeptide 9 (Cyp2b9) genes, arbitrarily selected from the microarray analysis, closely resemble the expressions determined by quantitative PCR. The data indicate that, E. papillata is associated with the induction of inflammatory response and with gene regulation in mice jejunum.

In vivo anticoccidial effects of Beta vulgaris (sugar beet) in broiler chickens.

Recently, use of botanicals as an alternative to anticoccidial drugs has been appealing approach for controlling avian coccidiosis. Therefore, this study was conducted to evaluate the anticoccidial activity of aqueous methanolic extract (100, 200 and 300 mg/kg of body weight) of Beta vulgaris (roots) in broiler chicks. A total of 315 day old broiler chicks were divided into seven equal groups (A, B, C, D, E, F and G). At 14th day of age, all groups except group G, which served as non infected non medicated control, were infected orally with 60,000 sporulated oocysts of mixed Eimeria species. At the same day, groups A, B and C were treated with graded oral doses of B. vulgaris aqueous methanolic extract (100, 200 and 300 mg/kg of body weight, respectively). Group D was treated with Vitamin-E, group E served as infected medicated control group (Baycox® treated) and group F served as infected non medicated control group (PBS treated). Treatment with extract, reference drug Baycox®, Vitamin E and PBS was continued for three consecutive days (14-16 days of age). Though, not at par with reference drug (Baycox®), B. vulgaris demonstrated good anticoccidial activity adjudged based on considered criteria, i.e., feed conversion ratio, lesion score, oocyst score and oocysts per gram of feces. Results of serum profile of infected chicks revealed no adverse effects of aqueous methanolic extract of B. vulgaris on the experimental chicks.

Prevalence of histopathological intestinal lesions and enteric pathogens in Dutch commercial broilers with time.

Intestinal disease has a major impact on the broiler industry due to economic and welfare reasons. Intestinal disease might occur due to a large number of reasons varying from well-defined pathogens to non-specific enteritis and complex syndromes. However, knowledge about the nature of intestinal disease and presence of enteric viruses in the Dutch broiler industry is largely absent. Therefore, a large-scale field study, in which 98 broiler flocks from 86 farms were sampled weekly, was started to assess the prevalence of histopathological lesions in the jejunum, a number of enterotropic viruses by real-time quantitative reverse transcriptase PCR (RT-qPCR) and coccidia by lesion scoring. Histopathological lesions indicative of intestinal disease were found in all flocks examined. The pathogens investigated were chicken astrovirus (99% of flocks positive), avian nephritis virus 3 (100%), rotavirus A (95%), rotavirus D (52%), reovirus (100%), Eimeria acervulina (94%), E. maxima (49%) and E. tenella (40%). The enteric viruses were more prevalent in the first weeks of the growing period, while coccidiosis was more frequently found at 4 and 5 weeks of age. The abundant presence of the enteric viruses and enteric disorders stresses the need to elucidate the role of these viruses in intestinal disease. Furthermore, the high prevalence of coccidiosis despite the use of anticoccidials shows that the current coccidial management programmes might be insufficient in controlling this disease.