Effect of Imidocarb dipropionate on the immune response to Foot and Mouth Disease vaccine in healthy and anaplasmosis-infected calves

Aim: This work was performed to investigate the effect of a potent anti-protozoan drug, Imidocarb on the cell mediated and humoral immune response to foot and mouth disease vaccine (FMDV, O strain) in normal and Anaplasmosis-infected calves. 1 Materials and Methods: A total of 55 male mixed bred calves were used and divided into two main groups of 25 calves each. The first group was healthy and the second was Anaplasma infected calves. FMDV was administered in both groups. Calves of the first and second groups were subdivided into equal five subgroups of 5 calves each. The first subgroup was vaccinated control. The treated subgroups were each given 3 mg / kg body weight Imidocarb dipropionate in a single intramuscular dose at one week before vaccination, at time of vaccination, one week and two weeks post vaccination with FMDV (O ), 1 respectively. The cellular immune response in the different groups was evaluated weekly, however antibody titers were measured by ELISA and serum neutralization test Results: Imidocarb increased rate of erythrocyte rosette forming lymphocytes when it was administered one week before vaccination, at time of vaccination and one week post vaccination. Imidocarb increased antibody titre of FMDV in both normal and anaplasmosis-infected calves. The protection rate due to challenge with virulent FMDV was high in treated calves as compared with the vaccinated control. Conclusion: The best immunopotentiating effect of Imidocarb is achieved by dosing one week before vaccinating calves with FMD vaccine.


Introduction
of cattle with babesiacides plays a central role in management of this disease [11].A wide range of Anaplasmosis is one of the most important tickantibacterial and antiprotozoal drugs have been used born disease (TBD) of cattle, worldwide [1,2,3].

Bovine anaplasmosis is caused by cattle infection with
Imidocarb dipropionate at the dose rate of 3mg/kg the tick-borne bacterium, Anaplasma marginale [4].
body weight is effective in prevention and treatment of Anaplasmosis, caused by the intraerythrocytic anaplasmosis in cattle [16].The infected animals were rickettsia Anaplasma marginale (Rickettsiales: treated with a single injection and carrier status was Anaplasmataceae), is an economically important successfully eliminated with two intramuscular disease of cattle which is endemic in tropical and injection of Imidocarb.Imidocarb is one of the most subtropical regions of the world [4][5][6][7].Infection of effective and successful in chemosterilization of cattle with A. marginale causes bovine anaplasmosis, a persistant Anaplasma marginale infection in cattle mild to severe hemolytic disease that results in [17].Moreover, the intraperitoneal administration of considerable economic loss to both dairy and beef Imidocarb significantly increased serum IL-10 levels industries [8] .This obligate intracellular pathogen can and lowered TNF-alpha levels in mice [18], suggesting be transmitted biologically by ticks, mechanically by that a novel anti-inflammatory effect of Imidocarb transfer of infective blood on the mouthparts of biting could be utilized to treat inflammatory conditions.The insects [4, 9], and, less commonly, by transplacental cell mediated immune response to FMD vaccine in transmission from dams to their calves [1].The method of ELISA was performed subgroup 1 were vaccinated with FMDV (O1 strain) according Ma et al. [22].and kept as nontreated control.The calves in subgroups 2, 3, 4, and 5 were given each 3 mg / kg body weight Challenge test: All calves at 21 days post vaccination Imidocarb dipropionate in a single intramuscular dose were challenged by FMDV; virulent strain and kept for at one week before vaccination, at the time of observation for 7 days.The protection percent was vaccination, one week post vaccination and two weeks calculated and recorded in all calves plus five post vaccination, respectively.The immune cellular susceptible control calves.response in the different groups against FMDV was Foot and Mouth Disease Virus (FMDV) "virulent evaluated weekly in normal calves for 8 weeks post strain": The calves were challenged with a FMDV, O / 1 vaccination.Humoral immunity was measured weekly 93 Aga virus (a stock glycerinated filtrate of bovine th for 9weeks then every 2 weeks up to the 17 week post tongue epithelium stored at -70°C).The virus was 4 vaccination.Blood Samples from the all groups (5 ml diluted to contain 10 mice protection ID50 per ml and each) were taken in sterilized heparinized centrifuge each calf was given 1ml by intradermolingual tube to be used for cell-mediated immune response; inoculation of 0.1 ml at ten sites.

Erythrocytes rosette per cent (ER %). Ten ml of blood
Statistical analysis: The statistical analysis was were collected in sterilized macartany bottle from all performed using the SPSS® 10.0 software package vaccinated animals for separation of serum to (SAS, Cary, NC, USA).Results are presented as determine the humoral immune response by ELISA arithmetic mean ± standard errors (SE).and serum neutralization test.

Effect of Imidocarb dipropionate on the cellular immune
The effect of Imidocarb dipropionate on cell response of calves was determined by measurement of mediated immunity of calves vaccinated with FMDV percent of Erythrocytes' Rosette forming lymphocytes was evaluated by measuring the affinity of T-(ER %).However, effect of Imidocarb on humoral lymphocytes to form a rosette with sheep RBCs immune response of vaccinated calves with FMDV (Table -1).There was a highly significant increase in the was evaluated by estimation of the antibody titre using rate of ER% in normal calves given Imidocarb before serum neutralization test (SN T), and ELISA test as and at the time of vaccination as compared with the well as challenge test.vaccinated control.However, the rate of ER% in Determination of percent of Erythrocytes Rosette Imidocarb was increased from the 2nd week after antibody titre when the drug was given one week treatment (Table-2).

anaplasmiod-infected calves and administered
before vaccination, and at the time of vaccination with Administration of Imidocarb at one week before FMDV.Moreover, the effect of Imidocarb dipropiovaccination increased the log titre of antibody by nate on the virus challenge test in normal and Anaplas-10 mosis-infected calves vaccinated with FMDV (O strain), using ELISA test in normal and anaplasmiod -infected 1 calves post vaccination with FMDV (Tables-3 and 4).
with 5 susceptible calves as a control for challenge.All When Imidocarb was given simultaneously with the vaccinated calves were protected; this was indicated vaccination and/or one week after vaccination, a by absence of tongue and foot lesions but the control significant increase in antibody titre was obtained from calves showed over 4 tongue lesions and lesions on the the second week post vaccination.Furthermore, 4 feet which indicate satisfactory challenge.Imidocarb administration at the time of and/ or one Discussion week post vaccination increased the antibody titre in Foot and Mouth Disease is an acute feverish and anaplasmiod-infected calves.The effect of Imidocarb contagious viral infection that can be seen in all on serum neutralization antibody titre in healthy and ruminating double nailed animals [23,24].FMD has anaplasmiod -infected calves and vaccinated with been known to be present in Egypt for more than 40 FMDV are shown in Tables-5 and 6.Intramuscular years causing drastic loses in milk and meat with deaths administration of Imidocarb dipropionate in a single among young animals [25].Theileriosis is a disease of dose of 3 mg/kg body weight increased the SN lymphoid organs of cattle resulting in profound leuco-treated groups injected before, simultaneously and/or penia, a high lymphoid cell parasiyosis and lymphocytes one week after vaccination, with a considerable increase lyses and thus reduction in the immunocompetence of in infected groups injected two weeks post vaccination.infected calves [17].The present study results show These results indicate that the drug may stimulate the that Imidocarb dipropionate in a dose of 3mg/kg b.wt.
humoral immune response against FMDV in both when injected intramuscularly in normal and anaplas-techniques (ELIZA and serum neutralization tests).miod-infected calves caused a significant increase in Moreover, the results of serological tests indicate that the rate of ER % in calves vaccinated with FMDV.vaccinated calves (normal and anaplasmiod-infected) These results suggested that the drug may stimulate the treated with Imidocarb gave rise to a high level of cellular immune response [2].A significant increase in antibodies which protected calves against subsequent the antibody titre against FMDV was recorded in the challenge.Similar results were described by The affinity of Twelfare and Institutional Animal Ethical Committee.lymphocytesto form a rosette with sheep RBCs was The animals were allocated to two groups of 25 calves each.Calves of the first group Therefore this study was aimed to investigate the approaches are needed [10].Vaccine use is not widely effect of Imidocarb on the cell-mediated and humoral available in several countries; consequently treatment immune response following foot and mouth disease vaccine in healthy and anaplasmosis -infected calves.according to the National regulations on animal forming lymphocytes (ER%): ® done according to the method of Kaupp et al. [20].Serum neutralization test: Neutralizing antibodies were free from Anaplasmosis and vaccinated with were detected by a microplate virus neutralization FMDV.The calves of the second group were assay modified by Pega et al. [21].Antibody tires were Anaplasma-infected animals and vaccinated with expressed as the reciprocal of the final dilution of FMDV.After that, each group were subdivided into serum in the serum/virus mixture.equal five subgroups of 5 calves each.The calves in ELISA test: