RNA Sequencing Analysis of Chicken Cecum Tissues Following Eimeria tenella Infection in Vivo.

Eimeria tenella (E. tenella) is one of the most frequent and pathogenic species of protozoan parasites of the genus Eimeria that exclusively occupies the cecum, exerting a high economic impact on the poultry industry. To investigate differentially expressed genes (DEGs) in the cecal tissue of Jinghai yellow chickens infected with E. tenella, the molecular response process, and the immune response mechanism during coccidial infection, RNA-seq was used to analyze the cecal tissues of an E. tenella infection group (JS) and an uninfected group (JC) on the seventh day post-infection. The DEGs were screened by functional and pathway enrichment analyses. The results indicated that there were 5477 DEGs (p-value < 0.05) between the JS and the JC groups, of which 2942 were upregulated, and 2535 were downregulated. GO analysis indicated that the top 30 significantly enriched GO terms mainly involved signal transduction, angiogenesis, inflammatory response, and blood vessel development. KEGG analysis revealed that the top significantly enriched signaling pathways included focal adhesion, extracellular matrix-receptor interaction, and peroxisome proliferator-activated receptor. The key DEGs in these pathways included ANGPTL4, ACSL5, VEGFC, MAPK10, and CD44. These genes play an important role in the infection of E. tenella. This study further enhances our understanding of the molecular mechanism of E. tenella infection in chickens.

Desempenho e estudo de depleção residual em frangos de corte infectados com eimerias e medicados com diclazuril / Performance and residual depletion study in chickens infected with eimerias and medicated with diclazuril

A avicultura brasileira atualmente ocupa o terceiro lugar, com uma produção anual de aproximadamente, 10,9 milhões de toneladas de carne de frango. Contudo, severas perdas econômicas são relatadas, devido à coccidiose em granjas de frangos de corte, matrizes e postura. As Eimerias são classificadas como protozoários, sendo que os mesmos se multiplicam nas células intestinais diminuindo a absorção de nutrientes, levando à desidratação, perda de sangue e susceptibilidade para infecção por outros micro-organismos. Com o desenvolvimento da pesquisa objetivou-se determinar os índices de produtividade zootécnica (ganho de peso, consumo de ração, conversão alimentar, taxa de mortalidade e índice de eficiência produtiva) bem como, mensurar o nível residual do Diclazuril nos tecidos de frangos de corte, comparando com os padrões internacionais de Limites Máximos de Resíduos determinados pelo Codex Alimentarius. Para a realização do estudo, utilizou-se 624 frangos de corte, onde metade do grupo foi inoculado experimentalmente com E. acervulina, E. maxima e E. tenella. O estudo foi composto por grupos tratados e não tratados com diclazuril. O uso do diclazuril expressou efeito positivo, no desempenho zootécnico das aves inoculadas artificialmente; a análise residual do medicamento apresentou um período de carência zero, sendo considerada segura para alimentação humana a carne de frangos medicados com Diclazuril.

Ação trófica de aditivos fitogênicos, glutamina e ácido glutâmico sobre a Bursa de Fabrícius e intestino delgado de frangos de corte / Trophic action of phytogenic additives, glutamine and glutamic acid on Bursa Fabricii and small intestine of broilers

The aim of this study was to evaluate the effect of phytogenic additives and glutamine plus glutamic acid, associated or not, on histomorphometry of bursa of Fabricius and small intestine, oocyst count and lesion scores, and carbon turnover of duodenal mucosa of broiler chickens infected with Eimeria acervulina. A total of 450 male broiler chickens was distributed into a completely randomized design with six treatments and three replications. Treatments consisted of control diet (CD); CD + coccidiosis vaccine; CD + antibiotic performance enhancers and anticoccidial (APE/AC); CD + glutamine and glutamic acid (Gln/Glu); CD + phytogenic additives (PA); CD + Gln/Glu + PA. Birds on treatment CD + vaccine were vaccinated via drinking water at three days of age against coccidiosis. At 16 days of age all birds of all treatments were inoculated orally and individually with 500,000 oocysts of Eimeria acervulina. There was no treatment effect on lesion score in the intestinal epithelium of birds. The smaller number of excreted oocysts was observed in groups of birds fed diets containing APE/AC and PA. Were observed better results of villus height and crypt depth for duodenum and ileum of birds of treatments containing Gln/Glu at 7 days of age, and Gln/Glu and PA at 21 days of age. Higher percentage of cortical area from bursa follicles was observed in birds fed diets supplemented with Gln/Glu and PA at 7, 14 and 21 days of age. Increased turnover of intestinal mucosa was observed in treatments containing Gln/Glu, indicating acceleration in development and regeneration of damaged tissue. Glutamine plus glutamic acid and phytogenic additives can provide improvements to structure, and thus to intestinal function, as well as to better immune response against the infectious challenges. Phytogenic additives can be used for coccidiosis control of broiler chickens where the use of antibiotic performance enhancers and anticoccidials is prohibited...

O objetivo deste trabalho foi avaliar o efeito dos aditivos fitogênicos e da glutamina mais ácido glutâmico, associados ou não, sobre a histomorfometria da Bursa de Fabricius e intestino delgado, sobre contagem de oocistos e escores de lesão e sobre o turnover do carbono da mucosa intestinal de frangos de corte experimentalmente infectadas com Eimeria acervulina. Para isso foram utilizados 450 pintos de corte machos distribuídos em delineamento inteiramente casualisado, com seis tratamentos e três repetições. Os tratamentos consistiram de dieta controle (DC); DC + Vacina de coccidiose; DC + antibióticos melhoradores de desempenho e anticoccidiano (AMD/AC); DC + glutamina e ácido glutâmico (Gln/Glu); DC + sditivos fitogênicos (AFs); DC + Gln/Glu + AFs. As aves do tratamento DC + Vacina foram vacinadas via água de bebida, aos três dias de idade, contra coccidiose. Aos 16 dias de idade todas as aves de todos os tratamentos foram inoculadas oralmente e individualmente com 500.000 oocistos de Eimeria acervulina. Não houve efeito dos tratamentos para escore de lesão no epitélio intestinal das aves. O menor número de oocistos excretados foi observado nos grupos de aves alimentadas com dieta contendo AMD/AC e AFs. Foram observados melhores resultados para altura das vilosidades e profundidade das criptas do duodeno e ílio das aves dos tratamentos contendo Gln/Glu, aos 7 dias de idade e Gln/Glu e AFs aos 21 dias de idade. Maior porcentagem de área cortical dos folículos bursais foi observada em aves alimentadas com dieta suplementada com Gln/Glu e AFs aos 7, 14 e 21 dias de idade. Maior turnover da mucosa intestinal foi observada em aves dos tratamentos contendo Gln/Glu, indicando aceleração do desenvolvimento e regeneração do tecido lesado. Glutamina mais ácido glutâmico e aditivos fitogênicos podem oferecer melhorias à estrutura e, consequentemente, à função do intestino, bem como melhores condições para resposta imune frente à desafios infecciosos...

Eficácia de Chenopodium ambrosioides (erva-de-santa-maria) no controle de endoparasitos de Coturnix japônica (codorna japonesa) / Effectiveness of Chenopodium ambrosioides (santa maria herb) for controlling Coturnix japonica (japanese quail) endoparasites

O objetivo foi testar in vitro e in vivo a eficácia da planta medicinal Chenopodium ambrosioidesLinnaeus, 1786 (erva-de-santa-maria), nas formas fitoterápica e homeopática, como meios alternativos para o controle de endoparasitos de Coturnix japonica Temminck & Schlegel, 1849 (codorna japonesa), um sério problema que afeta a criação e desempenho de aves domésticas, ocasionando morte quando muito intenso, retardo de crescimento, redução de índice de conversão alimentar e aumento na suscetibilidade às doenças infecciosas. As metodologias utilizadas foram preconizadas por Coles et al. (1992), creditada pela World Association for the Advancement of Veterinary Parasitology (WAAVP). A pesquisa evidenciou a presença dos gêneros Ascaridiae Eimeria. O ensaio in vitro demonstrou alta taxa de redução na inibição de eclosão de ovos de Ascaridiasp. (100,00%) e significativa taxa de redução na destruição de oocistos de Eimeriasp. (47,06%). O ensaio in vivodemonstrou alta taxa de redução na contagem de ovos de Ascaridiasp. nas fezes (100,00%) e expressiva taxa de redução na contagem de oocistos de Eimeriasp. nas fezes (60,33%). Chenopodium ambrosioides mostrou em certos momentos superioridade frente ao produto tradicional (Thiabendazole/Mebendazole) e índices superiores aos preconizados pelo Ministério da Agricultura do Brasil e Organização Mundial da Saúde como indicativos de eficácia.

The aim was in vitro and in vivo to test the effectiveness of Chenopodium ambrosioides Linnaeus, 1786 (santa maria herb) medicinal plant, in regard to phytotherapeutic and homeopathic forms as alternative methods to control Coturnix japonica Temminck & Schlegel, 1849 (japanese quail) endoparasites. The parasitosis is a serious problem affecting domestic poultry raising and performance causing death, delay in grow, food conversion rate reduction and increase of susceptibility to infectious diseases. Methodologies were advocated by Coles et al. (1992), corroborated by World Association for the Advancement of Veterinary Parasitology (WAAVP). Presence of the genera Ascaridia and Eimeria was displayed by this survey. In vitro essay demonstrated high reduction rate on eggs eclosion inhibition of Ascaridia sp. (100.00%) and significant reduction rate on oocyst destruction of Eimeria sp. (47.06%). In vivo essay demonstrated high fecal egg counting reduction rate of Ascaridia sp. (100.00%) and expressive fecal oocyst counting reduction rate of Eimeriasp. (60.33%). C. ambrosioides showed upper rates front traditional product (Thiabendalol/Mebendazol) as well as to those ones advocated by the Brazilian Ministry of Agriculturel and the World Health Organization as effectiveness indicative.

Occurrences of Eimeria spp. and gastrointestinal nematodes in dairy calves in southern Minas Gerais, Brazil / Ocorrência de Eimeria spp. e nematódeos gastrintestinais em bezerros no sul de Minas Gerais, Brasil

The aim of this cross-sectional observational study was to determine the frequency and factors associated with infection by Eimeria spp. and gastrointestinal nematodes in 356 calves on 20 dairy farms located in southern Minas Gerais, Brazil. Ten species of Eimeria spp. were identified, of which E. bovis (37.6%) and E. zuernii (17.9%) were the most frequent. From fecal cultures, four genera of gastrointestinal nematodes were recovered, of which Cooperia spp. (74.6%) and Haemonchus (19.4%) were the most frequent. Variables relating to higher levels of technology used on dairy farms showed a significant association (p < 0.05) with higher OPG and EPG counts, and are discussed in this study.

O objetivo deste estudo foi determinar a frequência e os fatores associados à infecção por Eimeria spp. e nematódeos gastrintestinais, em 356 bezerras provenientes de 20 rebanhos leiteiros, localizados no sul de Minas Gerais, Brasil. Foram identificadas dez espécies de Eimeria spp., sendo E. bovis (37,6%) e E. zuernii (17,9%) as mais frequentes. Nas coproculturas, foram recuperados quatro gêneros de nematódeos gastrintestinais, sendo os mais frequentes Cooperia spp. (74,6%) e Haemonchus spp. (19,4%).Variáveis relacionadas a um maior nível de tecnificação das propriedades leiteiras apresentaram associação significativa (p < 0,05), com maiores contagens de OoPG e OPG, e são discutidas neste estudo.

Cross-reactions between Eimeria falciformis and Eimeria pragensis in mice induced by trickle infections.

We describe a laboratory model using Eimeria falciformis and E. pragensis to investigate some of the interactions in double-species infections of Eimeria. Mice were given trickle infections by oral inoculation of 100 sporulated oocysts of one species at 3 or 4 day intervals throughout the experiments and, once immunity had developed, as indicated by cessation of oocyst production, the animals were challenged with a single inoculation of the other species. A trickle infection of E. falciformis gave a significant enhancement of oocyst output from E. pragensis infection as compared with animals that had not received E. falciformis. Histological examination of the mice infected with E. pragensis showed significantly more asexual parasites and a significantly higher female/male ratio in the mice that had received trickle E. falciformis infections than in those that had not. There was no evidence to suggest that extra asexual generations were occurring in these mice. In the converse experimental protocol, trickle E. pragnesis infections had no significant effect on a single E. falciformis infection. We discuss the possible mechanisms for the interactions and also how these interactions may influence multiple species infections in animals in their natural habitats.

Eimeria minasensis n.sp. (Apicomplexa: Eimeriidae) in the domestic goat Capra hircus, from Brazil

Eimeria minasensis n.sp. is described in the domestic goat Capra hircus from Brazil. Oocysts ellipsoidal are 35 x 24.5 (32-37.7 x 20.9-27.9) µm. Sporocysts elongate-ellipsoid are 15.2 x 9 (12.3-18.4 x 7.8-10.2) µm, with a Stieda body at the narrow end. Oocyst wall smooth and bilayered; outer layer about 1.2 (0.8-1.6) µm and colorless; inner layer about 0.5 (0.4-0.8) µm and dark-brown. Micropyle, a mound-shaped micropylar cap 1,6 x 8,9 (0,8-2 x 7-10,2) easily dislodged; one or more oocyst polar granules present. Oocyst residuum absent. Sporocyst residuum present, composed of many scattered granules. Sporozoites elongate, lying lengthwise, "head to tail" in the sporocysts; one or two refractile globules are usually visible. Sporulation time was 120 hr at 27ºC, prepatent period, 19 to 20 days and patent period 15 to 25 days. Gamonts, gametes and oocysts present in cecum and colon. Prevalence was 12.8 per cent (6/47) in goats from Minas Gerais, Brazil.

Passive immunization against somatostatin increases resistance to Eimeria vermiformis infection in susceptible mice.

The effect of in vivo immunoneutralization of somatostatin (SRIF) on Eimeria vermiformis intestinal infection was studied in resistant (BALB/c), and susceptible (C57BL/6) mouse strains. An anti-SRIF monoclonal antibody (MAb-SRIF) was used to passively immunize the mice by intraperitoneal injection. The animals were subsequently orally infected with oocysts of E. vermiformis. Individual fecal samples were collected daily for 21 days to monitor the kinetics of oocyst shedding. The fecal oocyst shedding was significantly higher in the C57BL/6 strain than in the BALB/c strain (P < 0.01). Passive immunization with MAb-SRIF in the C57BL/6 mice significantly reduced the number of oocysts in feces (P < 0.05), when compared to the infected non-immunized mice of the same strain. Infected BALB/c mice showed no difference in oocyst shedding in response to the passive immunoneutralization with MAb-SRIF. In conclusion, passive immunization with MAb-SRIF increased resistance to E. vermiformis-infection in the susceptible C57BL/6 mice, but not in the resistant BALB/c mice. This suggests that SRIF modulates gut immune function in parasitic infection.

[Role of flies in the epizootiology of coccidiosis in poultry]. / Vurkhu roliata na mukhite v epizootologiiata na koktsidiozata po ptitsite.

The possibility is shown Musca domestica L. to ingest considerable amounts of oocysts of Eimeria tenella which remain in their digestive tract during 120 hrs. Both sporulated and non-sporulated oocysts remain viable and infectable. Non-sporulated oocysts do not form spores in the digestive tracts of flies, but after their excretion, they terminate normally their development under favourable conditions. It is shown that when flies, infected with oocysts, are fed to susceptible chicks, they are affected with coccidiosis.

A fine structural study of asexual stages of the murine coccidium Eimeria ferrisi Levine and Ivens 1965.

The schizogony of Eimeria ferrisi was studied in experimentally infected Mus musculus. Developmental stages occurred in epithelial cells of the cecum and colon. During transformation of invasive stages into schizonts the inner membrane complex of the pellicle, the conoid, subpellicular microtubules and micronemes gradually disappeared. The micropore, however, seemed to persist. Dividing nuclei had eccentric intranuclear spindles consisting of microtubules which extended between 2 centrocones, in close relationship with centrioles. During the last nuclear division anlagen of merozoites appeared as extensions on the surface of schizonts. The outer single membrane of the schizont became the outer membrane of the merozoite pellicle. Cytoplasmic organelles, typical of eimerian merozoites were incorporated into the developing merozoites. Finally the merozoite became detached leaving behind a residual cytoplasm. Fully developed merozoites had a 3-layered pellicle, the outer single unit membrane was continuous around the merozoite with the inner complex having interruptions at the anterior and posterior poles and at the micropores. Thirty-two subpellicular microtubules, originating at the anterior polar ring extended to the posterior region of each merozoite. The apical complex consisted of a conoid, preceded by 2 rings and surrounded by a polar ring. Two rhoptries were present having club-shaped terminal ends and slender ductules in the conoid region. Some merozoites had enlarged rhoptries, with the distal vesical appearing dense and osmiophilic. The Golgi complex, endoplasmic reticulum, mitochondria, polysaccharide granules were similar to those seen in other eimerian merozoites.

Structure and mode of function of the organelles associated with nutrition of the macrogametes of Eimeria acervulina.

The macrogamete of Eimeria acervulina, lay and developed within the host cell in a parasitophorous vacuole. The cytoplasmic membrane of the host cell bordering the vacuole was not smooth, but it had numerous folds extending into the vacuole. These "intravacuolar folds" varied in depth and number in different sections. In some, the majority of the folds were disconnected from the host cell. Once disconnected, they evidently disintegrated forming the amorphous, particulate material present in the parasitophorous vacuole. The pellicle of the young macrogamete consisted of a single unit membrane with an osmiophilic material representing the second membrane. Two unit membranes were apparent at a later stage of development when the wall-forming bodies had been formed and amylopectin granules deposited. Two kinds of organelles were present on the surface of the macrogamete, typical micropores and invaginations of the pellicle. The micropores arose from an invagination of the outer membrane, which continued through the invagination without interruption. Irrespective of whether an inner membrane was present in the pellicle or not, a thickened cylindrical wall around the inner portion of the invagination was always present. Micropores appeared in large numbers in both micro- and macrogametocytes. As many as three micropores were seen in a surface area of 2 mu2. Invaginations arose in a similar manner by infolding of the pellicle. They differed from micropores in that the thickened cylindrical wall present around the inner portion of the micropore was absent, and also in that invaginations had no uniform appearance. They were of varying shapes, and lengths, varying from very short V-shaped to long and narrow. Micropores and invaginations take in nutrients in the form of particulate matter present in the parasitophorous vacuole, this material having been derived from the host-cell membranous "intravacuolar folds". The micropores function as cytostomes and the invaginations take in material by means of pinocytosis. Large numbers of intravacuolar tubules were seen at the surface of the macrogamete. They were present only at certain areas of the macrogamete and in groups and were connecting the parasite with the host cell. They were about 80-110 nm in diameter, and were seen to attain a length of up to 6 mu. Evidence was obtained indicating that the tubules transport free ribosomes from the host cell to the parasite. The ribosomes were seen to accumulate in "pockets" within the cytoplasm of the host cell, at the area where the tubules were connected.