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Background: Coccidiosis is an important parasitic disease of chickens, causing high mortality and morbidity. This morbidity is believed to be correlated with altered population dynamics of blood cells and immunocompromisation. Objectives: This study investigated the effects of mixed Eimeria species (viz., tenella, maxima, acervulina and necatrix) infection on hematology and immune responses following Newcastle disease (ND) and infectious bursal disease (IBD) booster vaccination in broilers. Animals and methods: One-day-old broiler chicks (Hubbard; n D 200) were divided into two equal groups A and B. On day 16, group A was infected orally with Eimeria species (7 £ 10 4 sporulated oocysts), whereas group B served as control. Both groups were analyzed for hematological parameters on post-infection days 6À8. Sera from both groups were analyzed for antibody titers against ND and IBD vaccines. On day 8 post-infection, lymphoid organs were also examined. Results: Significantly lower (P < 0.05) levels of plasma proteins, globular volume, hemoglobin concentration, packed cell volume, total erythrocytes, mean corpuscular volume and mean corpuscular hemoglobin concentration were found in infected chickens compared with non-infected control chickens. In addition, the infected group exhibited significantly increased (P < 0.05) numbers of different leukocytes. Infected chickens also showed significantly lower antibody titers against ND and IBD with decreased relative organ weights of all lymphoid organs except spleen. Conclusion and recommendations: Mixed species of Eimeria adversely affected the hematology and immune efficiency of broilers. Thus, inexpensive immune potentiators and hemotonics along with appropriate anti-coccidial medications are suggested to avoid the complications and subsequent economic losses.

The present study reports the effect of Emblica officinalis (EO) derived tannins on humoral immune responses and their protective efficacy against Eimeria infection in chickens. Tannins were extracted from EO and characterized by HPLC. EO derived tannins (EOT) and commercial tannins (CT) were orally administered in broiler chicks in graded doses for three consecutive days, that is, 5th-7th days of age. On day 14 after administration of tannins, humoral immune response was detected against sheep red blood cells (SRBCs) by haemagglutination assay. Protective efficacy of tannins was measured against coccidial infection, induced by Eimeria species. Results revealed higher geomean titers against SRBCs in chickens administered with EOT as compared to those administered with CT and control group. Mean oocysts per gram of droppings were significantly lower (P < 0.05) in EOT administered chickens as compared to control group. Lesion scoring also showed the lowest caecal and intestinal lesion score of mild to moderate intensity in chickens administered with EOT. Further, significantly higher (P < 0.05) daily body weight gains and antibody titers were detected in EOT administered chickens as compared to those of CT administered and control groups. EOT showed the immunostimulatory properties in broilers and their administration in chickens boost the protective immunity against coccidiosis.

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Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are the most pathogenic and economically important mycoplasma pathogens that infect chickens. The development of rapid and innovative molecular diagnostic techniques is of pivotal importance for their effective control. The aim of the present study was to develop a novel duplex TaqMan real-time PCR assay for the simultaneous detection of MG and MS. This duplex real-time PCR assay incorporates TaqMan (FAM/NED) labelled minor groove binder (MGB) probes that target the cytadhesin encoding surface protein (mgc2) gene and the haemagglutinin surface protein (vlhA) gene of MG and MS, respectively. The assay also contained a TaqMan exogenous internal positive control (Exo IPC), to avoid false negative results that might happen due to failure in DNA extraction/PCR inhibition. The TaqMan MGB probe-based duplex RT-PCR incorporating Exo IPC was then applied to DNA from culture isolates for the simultaneous detection of MG (mgc2 gene) and MS (vlhA gene). For duplex RT-PCR the sensitivity recorded was 10 -3 CFU/ml and 10 -2 CFU/ml for MG and MS template DNA, respectively. The specificity of the real-time PCR assay was 100% for MG-and MS-specific probes in detecting both single as well as double infections. In conclusion, the use of TaqMan MGB probes for the detection of mgc2 and vlhA genes confers extra specificity and the incorporation of Exo IPC simplifies the assay, allowing the detection of double infections with low-copy target DNAs.Keywords: avian Mycoplasma spp.; duplex real-time PCR; MGB probe; mgc2 gene; vlhA gene; internal positive control Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are the most pathogenic and economically significant mycoplasma pathogens that infect chickens. The three major approaches currently used for the diagnosis of avian mycoplasmas are isolation and identification, detection of antibodies and molecular detection of the organisms by polymerase chain reaction (Raviv and Kleven 2009). PCR based on detection of mgc2-cytadhesin encoding surface protein gene for MG (Garcia et al. 2005), and vlhA-haemagglutinin surface protein gene of M. synoviae (Hong et al. 2004) are the most widely used methods for the detection, typing and determination of the source of infection. Real-time PCR (RT-PCR) avoids the need for post amplification processing, which saves time and labour compared to conventional PCR methods (Kawahara et al. 2008). TaqMan RT-PCR is a simpler method on which to base a multiplex PCR assay to detect mixed infections in a single PCR reaction. So far, only a limited number of TaqMan based diagnostic RT-PCR assays have been reported that target the mgc2 gene (Grodio et al. 2008;Raviv and Kleven 2009;Sprygin et al. 2010) for detection of MG, and the vlhA gene for detection of MS. The duplex RT-PCR-based detection of MG and MS using the 269Veterinarni Medicina, 60, 2015 (5): 268-273 Original Paper (OIE 2008). The avian mycoplasma RT-PCR assays previously described by Grodio et al. (2008) and Sprygin et al. ...