In vitro and in vivo evaluation of chlorhexidine salts as potential alternatives to potassium dichromate for Eimeria maxima M6 oocyst preservation.

Introduction: Coccidiosis caused by the Eimeria spp., an Apicomplexan protozoon, is a major intestinal disease that affects the poultry industry. Although most cases of coccidiosis are subclinical, Eimeria infections impair bird health and decrease overall performance, which can result in compromised welfare and major economic losses. Viable sporulated Eimeria oocysts are required for challenge studies and live coccidiosis vaccines. Potassium dichromate (PDC) is typically used as a preservative for these stocks during storage. Although effective and inexpensive, PDC is also toxic and carcinogenic. Chlorhexidine (CHX) salts may be a possible alternative, as this is a widely used disinfectant with less toxicity and no known carcinogenic associations. Methods: In vitro testing of CHX gluconate and CHX digluconate exhibited comparable oocyst integrity and viability maintenance with equivalent bacteriostatic and bactericidal activity to PDC. Subsequent use of CHX gluconate or digluconate-preserved Eimeria oocysts, cold-stored at 4°C for 5 months, as the inoculum also resulted in similar oocyst shedding and recovery rates when compared to PDC-preserved oocysts. Results and discussion: These data show that using 0.20% CHX gluconate could be a suitable replacement for PDC. Additionally, autofluorescence was used as a method to evaluate oocyst viability. Administration of artificially aged oocysts exhibiting >99% autofluorescence from each preserved treatment resulted in no oocyst output for CHX salt groups.

Protective Immunity Induced by an Eimeria tenella Whole Sporozoite Vaccine Elicits Specific B-Cell Antigens.

This study investigated protection against Eimeria tenella following the vaccination of chicks with 5.3 × 106 E. tenella whole-sporozoites emulsified in the nanoparticle adjuvant IMS 1313 N VG Montanide™ (EtSz-IMS1313). One-day-old specific pathogen-free (SPF) chicks were subcutaneously injected in the neck with EtSz-IMS1313 on the 1st and 10th days of age. Acquired immunity was assayed through a challenge with 3 × 104 homologous sporulated oocysts at 21 days of age. The anticoccidial index (ACI) calculated for every group showed the effectiveness of EtSz-IMS1313 as a vaccine with an ACI of 186; the mock-injected control showed an ACI of 18 and the unimmunized, challenged control showed an ACI of -28. In a comparison assay, antibodies from rabbits and SPF birds immunized with EtSz-IMS1313 recognized almost the same polypeptides in the blotting of E. tenella sporozoites and merozoites. However, rabbit antisera showed the clearest recognition pattern. Polypeptides of 120, 105, 94, 70, 38, and 19 kDa from both E. tenella life cycle stages were the most strongly recognized by both animal species. The E. tenella zoite-specific IgG antibodies from the rabbits demonstrated the feasibility for successful B cell antigen identification.

Hallazgo de Eimeria tenella en células epiteliales de la bolsa de Fabricio / Findings of Eimeria tenella in epithelial cells of the bursa of Fabricius

En el presente trabajo se describen observaciones realizadas durante un experimento diseñado para estudiar el comportamiento de los leucocitos polimorfonucleares en infecciones con E. tenella en pollos tratados con 5-FU como agente granulocitopénico. Ciento veinte pollitos de engorda fueron asignados en 4 grupos con 30 pollitos cada uno: 1) testigo blanco, 2) tratado con 200 mg/kg de peso de 5-fluorouracilo (5-FU), 3) infectado oralmente con 500 ooquistes esporulados de Eimera tenella y 4) infectado con E tenella después del tratamiento con 5-FU. Se administró E. tenella a 5 pollos de cada grupo a los días: 2, 4, 6, 8, 10 y 12 días posinoculación con 5-FU (17 días de edad). Siete días posinfección con E. Tenella se tomaron muestras de tejidos de varios órganos, para su estudio histológico. Se observaron coccidias en diferentes estado de desarrollo en 25 y 24 pollos en los grupos 3 y 4, respectivamente. Se observó hiperplasia epitelial moderada a severa, infiltrado mononuclear en el subepitelio, y algunos quistes intraepiteliales, que fueron asociados con la presencia del parásito. Este hallazgo muestra que bajo ciertas circunstancias E. tenella es capaz no sólo de invadir el epitelio de la bolsa de Fabricio, sino tambien de desarrollarse en él