Systemic inflammatory response indicators in rabbits (Oryctolagus cuniculus) experimentally infected with sporulated oocysts of Eimeria stiedai (Apicomplexa: Eimeriidae)

Hemograms and acute-phase proteins in adult male New Zealand White rabbits that had been experimentally infected orally with sporulated oocysts of Eimeria stiedai were evaluated over a 28-day period. Fifty animals were used, divided into two groups: group A infected with 1 x 10(4) sporulated oocysts of E. stiedai and group B inoculated with distilled water. on the seventh day after infection, the infected animals presented anemia and leukocytosis with neutrophilia and monocytosis. Protein fractionation by means of electrophoresis identified 19 acute-phase proteins with molecular weights ranging from 24 to 238 kD. Ceruloplasmin, transferrin and haptoglobin showed high levels on the seventh day after infection, with gradual increases in their concentrations until the end of the experimental period. Thus, from the data of the present study, E. stiedai is considered to be a pyogenic etiological agent for which the infection level can be monitored through the leukocyte count and serum concentrations of ceruloplasmin, transferrin and haptoglobin, and these can be recommended as complementary tests.

Eimeria stiedai: Metabolism of lipids, proteins and glucose in experimentally infected rabbits, Oryctolagus cuniculus

Rabbits were experimentally infected with sporulated Eimeria stiedai oocysts. A total of 50 white adult rabbits, New Zealand race, were distributed into two groups: Group A was infected with 1x10 4 sporulated Eimeria stiedai oocysts, while group B was inoculated with distilled water as a control. The animals generally displayed increased levels of total protein, globulin, total cholesterol, LDL-c and triacylglycerols; however, total levels of liver lipids and HDL-c decreased, and plasma glucose levels varied during the experimental period. In sum, Eimeria stiedai infection of rabbits caused a considerable number of changes in the metabolism of lipids, proteins and glucose, which is likely due to direct effects of liver cirrhosis on normal body function.

Lipid and glucose metabolism of broilers (Gallus gallus domesticus) experimentally infected with Eimeria acervulina Tyzzer, 1929 oocysts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aspectos clínicos e patológicos em frangos de corte (Gallus gallus domesticus) infectados experimentalmente com oocistos esporulados de Eimeria acervulina Tyzzer, 1929

No presente estudo, sinais clínicos e alterações patológicas foram avaliados por 30 dias em frangos de corte, linhagem Cobb, machos, com dez dias de idade, infectados com Eimeria acervulina. Foram utilizados 192 animais distribuídos em 3 grupos: grupo A inoculado com 1x10(6) oocistos esporulados; grupo B inoculado com 1x10(5) oocistos esporulados; grupo C inoculado com água destilada. Os sinais clínicos observados foram anorexia, diarréia e apatia. As alterações patológicas macroscópicas observadas foram: enterite, hiperemia seguido de congestão intestinal, excesso de exsudato mucoso no lúmen do intestino delgado, palidez e desidratação muscular, alto acúmulo de bile na vesícula biliar e deposição de gordura hepática. A atrofia de vilosidades e alta presença de células inflamatórias foram as alterações microscópicas observadas no epitélio intestinal. Na análise histopatológica do fígado observaram-se infiltrados inflamatórios e deposição de gordura. Os resultados demonstraram que frangos de corte infectados experimentalmente com E. acervulina apresentam progressivas lesões intestinais de intensidade variável e que essas anormalidades são as principais causas de redução no desenvolvimento da ave.

Molecular Identification and Characterization of Ileal and Cecal Fungus Communities in Broilers Given Probiotics, Specific Essential Oil Blends, and Under Mixed Eimeria Infection

Broiler digestive tract fungal communities have gained far less scrutiny than that given corresponding bacterial communities. Attention given poultry-associated fungi have focused primarily on feed-associated toxin-producers, yeast, and yeast products. The current project focused on the use of pyrosequencing and denaturing gradient gel electrophoresis (DGGE) to identify and monitor broiler digestive fungal communities. Eight different treatments were included. Four controls were an Uninfected-Unmedicated Control, an Unmedicated-Infected Control, the antibiotic bacitracin methylene disalicylate plus the ionophore monensin as Positive Control, and the ionophore monensin alone as a Negative Control. Four treatments were two probiotics (BC-30 and Calsporin) and two specific essential oil blends (Crina Poultry Plus and Crina Poultry AF). All chickens except the Unmedicated-Uninfected Control were given, at 15 days of age, a standard oral Eimeria inoculum of sporulated oocysts. Ileal and cecal digesta were collected at pre-Eimeria infection at 14 days of age and at 7 days post-Eimeria infection at 22 days of age. Extracted cecal DNA was analyzed by pyrosequencing to examine the impact of diet supplements and Eimeria infection on individual constituents in the fungal community, while DGGE was used to compare more qualitative changes in ileal and cecal communities. Pyrosequencing identified three phyla, seven classes, eight orders, 13 families, 17 genera, and 23 fungal species. Ileal and cecal DGGE patterns showed fungal communities were clustered mainly into pre- and post-infection patterns. Post-infection Unmedicated-Uninfected patterns were clustered with pre-infection groups demonstrating a strong effect of Eimeria infection on digestive fungal populations. These combined techniques offered added versatility towards unraveling the effects of enteropathogen infection and performance enhancing feed additives on broiler digestive microflora.

Eimeria Spp. in Brazilian Water Buffalo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identification of Eimeria mitis and Eimeria praecox in broiler feces using polymerase chain reaction

There are few reports concerning the epidemiology of Eimeria praecox and Eimeria mitis in Brazil. In the present experiment, the polymerase chain reaction (PCR) was used to identify these species in 156 samples of broiler chicken feces from several Brazilian states and the Federal District. Oocysts present in feces samples were purified by sodium chloride flotation followed by addition of DNAzol reagent (Invitrogen®) for extraction of genomic DNA. DNA was precipitated and stored following DNAzol reagent manufacture's instructions. The primers and PCR conditions were as described by Schnitzler et al. (1999). In the 156 field samples analyzed by PCR, 70 and 45 were positive for E. praecox and E. mitis, respectively. In this study we have shown that DNA extraction using DNAzol followed by PCR can be a useful tool in epidemiological studies, since it provides fast and reliable detection of Eimeria sp. in field samples.

Anticoccidial effects of coumestans from Eclipta alba for sustainable control of Eimeria tenella parasitosis in poultry production

The aim of this study was to evaluate the anticoccidial efficacy of a product containing coumestans from Eclipta alba. Experimental conditions were set up as to reproduce the environment conditions for husbandry adopted in commercial broiler farms. Broilers were raised in broiler chicken shed provided with feeders, drinkers, illumination and temperature control systems and floor covering to afford an adequate nourishing environment. Male Cobb broilers (240) were assigned to four experimental groups being each experimental group set apart in rice straw-covered shed isolated with wire mesh. One-day-old broilers were reared in a coccidian-free environment with ad libitum supply of filtered water and freely available standard feed, from the 1st to the 35th day of life. The T1 group received standard feed (negative control); T2 was treated with standard feed supplemented with 66 ppm of salinomycin (positive control); groups T3 and T4 had standard feed supplemented with the ethyl acetate fraction from methanolic extract of E. alba aerial parts, which contains the coumestans WL and DWL (120 and 180 ppm, respectively). The chicken broilers were individually infected with 2 x 104 oocysts of Eimeria tenella when they were 14 days old and were monitored weekly to evaluate zootechnical parameters such as weight gain and food conversion ratio. Counting of coccidial oocyst in chiken feces was assessed from random samples, from the 21st to 28th days of life, which corresponded to 7-14 days after the infection. Five chickens selected at random from each experimental group were subsequently euthanized at 21, 28 or 35 days of life to determine the lesion score in the cecal region and to excise a cecum portion for histopathological evaluation. The group treated with coumestans from E. alba presented an average weight gain and food conversion ratio higher than the negative control group and similar to the mean value of the positive control group. Coumestan-treated groups showed a significant decrease in the oocyst counting since the 21th day of life and displayed a reduced number of macroscopic lesions. Histopathological evaluations of cecum fragments showed that both treatments induced the migration of defense cells at the site of infection. A severe destruction of the cecal lining was found in the intestinal tract of broilers fed with a coumestans dose of 180 ppm. Overall, our results validate the use of a phytotherapy containing E. alba coumestans at a dose of 120 ppm as a therapeutic or prophylactic agent against avian coccidiosis. (C) 2010 Elsevier B.V. All rights reserved.

Use of Pyrosequencing and Denaturing Gradient Gel Electrophoresis to Examine the Effects of Probiotics and Essential Oil Blends on Digestive Microflora in Broilers Under Mixed Eimeria Infection

A protective digestive microflora helps prevent and reduce broiler infection and colonization by enteropathogens. In the current experiment, broilers fed diets supplemented with probiotics and essential oil (EO) blends were infected with a standard mixed Eimeria spp. to determine effects of performance enhancers on ileal and cecal microbial communities (MCs). Eight treatment groups included four controls (uninfected-unmedicated [UU], unmedicated-infected, the antibiotic BMD plus the ionophore Coban as positive control, and the ionophore as negative control), and four treatments (probiotics BC-30 and Calsporin; and EO, Crina Poultry Plus, and Crina PoultryAF). Day-old broilers were raised to 14 days in floor pens on used litter and then were moved to Petersime batteries and inoculated at 15 days with mixed Eimeria spp. Ileal and cecal samples were collected at 14 days and 7 days postinfection. Digesta DNA was subjected to pyrosequencing for sequencing of individual cecal bacteria and denaturing gradient gel electrophoresis (DGGE) for determination of changes in ileal and cecal MC according to percentage similarity coefficient (%SC). Pyrosequencing is very sensitive detecting shifts in individual bacterial sequences, whereas DGGE is able to detect gross shifts in entire MC. These combined techniques offer versatility toward identifying feed additive and mild Eimeria infection modulation of broiler MC. Pyrosequencing detected 147 bacterial species sequences. Additionally, pyrosequencing revealed the presence of relatively low levels of the potential human enteropathogens Campylobacter sp. and four Shigella spp. as well as the potential poultry pathogen Clostridiun perfringens. Pre- and postinfection changes in ileal (56%SC) and cecal (78.5%SC) DGGE profiles resulted from the coccidia infection and with increased broiler age. Probiotics and EO changed MC from those seen in UU ilea and ceca. Results potentially reflect the performance enhancement above expectations in comparison to broilers not given the probiotics or the specific EO blends as feed supplements.

Espécies do gênero Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) em caprinos leitros mantidos em sistema intensivo na região de São Jose do Rio Preto, estado de São Paulo, Brasil.

Were examined 58 dairy goats, 41 kids and 17 adults, Saanen and Alpine breeds, males and females, in intensive system, in the São José do Rio Preto region, São Paulo state, for detection and identification of Eimeria spp. oocysts. The 58 feces samples analyzed by the Centrifugal-flotation technique, were positive for, at least, one Eimeria species. The Eimeria species found in this research were: E. ninakohlyakimovae (77,6%), E. jolchijevi (72,4%), E. alijevi (63,8%), E. christenseni (63,8%), E. arloingi (62,1%), E. caprovina (56,9%), E. hirci (50,0%) and E. caprina (48,3%). It follows that the high rate of the positive animals and the high frequency of the Eimeria species among the animals demonstrated that the disease is common in dairy goats, kids and adults, in intensive system.

Espécies do gênero Eimeria (Apicomplexa: Eimeriidae) em Tamanduás-bandeira (Myrmecophaga tridactyla Linnaeus, 1758) em cativeiro.

The parasitism of the two giant anteaters adults (Myrmecophaga tridactyla), one male and one female, infected naturally with Eimeria escomeli, E. tamanduae e E. marajoensis was related in the present research. In E. escomeli oocysts were 23.9 +/- 1.89 by 19.7 +/- 1.60 microm and its sporocysts were 11.47 +/- 1.25 by 6.48 +/- 0.80 microm. In E. tamanduae oocysts were 23.52 +/- 0.95 by 20.59 +/- 0.92 microm and its sporocysts were 12.19 +/- 0.65 by 7.15 +/- 0.55 microm. In E. marajoensis oocysts were 13.5 +/- 1.7 by 13.1 +/- 1.8 microm and its sporocysts were 7.4 +/- 0.58 by 5.4 +/- 0.8 microm. Eimeria escomeli was described before parasitizing giants anteater from Bolivia, and it was point out as the first time in Brazil. The presence of E. tamanduae and E. marajoensis parasitizing giant anteaters indicate the possibility of having co-infection of them among animals of the family Myrmecophagidae.

Um surto de coccidiose em perdizes (Rhynchotus rufescens), criadas em cativeiro, por Eimeria rhynchoti Reis e Nóbrega, 1936 (Apicomplexa: Eimeriidae).

Eimeria rhynchoti is redescribed parasitizing partridge (Rhynchotus rufescens), reared in captivity, from Jaboticabal City, São Paulo State, Brazil. Sporulation takes place in 48 hours, the shape of oocysts found vary from spherical to elliptic with 23.01 micro +/- 1.57 of length by 21.0 micro +/- 1.78 of width. The microple, polar cap and residuum of the oocysts were absent. The oocyst wall, measures 2.2 micro +/- 0.31 of thickness, is composed by two smooth layers; the polar granule is present. The sporocysts length was 15.03 mm +/- 2.12 by 8.08 mm +/- 0.84 of width vary from elliptic to elongate. Sporocyst wall slender with is fine and Stieda body; the residue found in form of several smaller granules spherical compacts. The sporozoites are contrary extending along the sporocysts wall possessing refracts body of easy visualization.

Avaliação fisiopatológica de coelhos (Oryctolagus cuniculus) infectados experimentalmente com oocistos esporulados de Eimeria stiedae (APICOMPLEXA: EIMERIIDAE)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Eimeria species in young and adult sheep raised under intensive and/or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil

The present study aimed to identify Eimeria species in young and adult sheep raised under intensive and / or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil using the traditional diagnostic methods and to correlate the infection level/types of infection in the different age/system in this herd. Fecal samples were collected from the rectum of 210 sheep and were subjected to laboratory analysis to differentiate the species. Furthermore, animals were observed to determine the occurrences of the clinical or subclinical forms of eimeriosis. Out of the 210 collected fecal samples, 147 (70%) were positive for Eimeria oocysts, and 101 (47.86%) belonged to young animals that were raised under intensive and / or semi-intensive farming systems. Oocysts from 9 species of Eimeria parasites were identified in the sheep at the following prevalence rates: E. crandallis, 50.0%; E. parva, 21.6%; E. faurei, 8.1%; E. ahsata, 8.1%; E. intricata, 5.4%; E. granulosa, 2.7%; E. ovinoidalis, 2.0%; E. ovina, 1.3%; and E. bakuensis, 0.6%. There were no differences regarding the more frequent Eimeria species among the different ages of animals or between the different farming management systems. Based on these data, E. crandallis was the most prevalent, followed by E. parva and E. faurei species, regardless of the age. Higher parasitism was diagnosed in the young animals that were raised in a confinement regime, and the disease found in the herd was classified as subclinical. Further studies should be conducted in this herd, to verify if the eimeriosis subclinical can cause damage especially in young animals with a high level of infection.

Efficacy of sulfadoxine plus trimethoprim compared to management measures for the control of Eimeria parasitism in naturally infected and clinically asymptomatic sheep that were maintained in a feedlot

The efficacy of sulfadoxine + trimethoprim in comparison to management measures for the control of Eimeria parasitism was studied in naturally infected sheep that were raised in a feedlot and were clinically asymptomatic for eimeriosis. Weight gain was also evaluated in these animals. The following groups were formed with 15 animals/group: TO!, control animals that received saline solution and maintenance of the same management measures that were performed before the study; T02, animals that received two intramuscular doses of sulfadoxine (20 mg/kg) + trimethoprim (4 mg/kg) with a 14-day interval; T03, sheep that received two intramuscular doses of sulfadoxine (20 mg/kg) + trimethoprim (4 mg/kg) with a 14-day interval plus management measures (wood shaving bedding was changed every Monday, and 30g of ammonium sulfate were applied to the bedding and other facilities were performed every Thursday, 10 mL/20 L of water); and T04, animals that received only the management measures described for the previous group. The highest efficacy rates (arithmetic mean) for the T02 group (sulfadoxine + trimethoprim at days 0 and 14) were 21.04% and 21.98% on the 14th and 28th days after the first treatment (DAFT), respectively. However, the treatment showed efficacy rates below 17% and was totally ineffective from the 70th DAFT to the end of the study. In both the T03 (chemical treatment+ management) and T04 (management only) groups, a significant (P <= 0.05) reduction of oocyst shedding per gram of feces was observed in the animals from the 14th DAFT in comparison to the control group; however, an efficacy rate above 90% was observed from the 28th DAFT. Animals belonging to the T02, T03 and T04 groups presented with alterations in weight gain of 0.57 kg, 4.30 kg and 4.53 kg, respectively, in comparison with the control animals (T01) throughout the 91-day study period. Thus, it is possible to conclude that the two-dose sulfadoxine + trimethoprim treatment, given with a 14-day interval, had little no effect on the oocyst shedding. Moreover, the adopted management measures were enough to cause a significant decrease in the animal parasite loads. (C) 2013 Published by Elsevier B.V.