In nature, herpesviruses are mostly associated with a single host species. Italian Mediterranean buffaloes (Bubalus bubalis) is associated with Bubaline herpesvirus 1 (BuHV-1) but also with Bovine herpesvirus 1 (BoHV-1). Like different alphaherpesvirinae subfamily members, the primary site for BuHV-1 or BoHV-1 latency is sensory neurons within trigeminal ganglia (TG). Recently, it has been shown that cattle infected by BoHV-1 seemed to have more oxidative stress and low antioxidant defense (Durgut et al., 2013). Thus, to better understand the physiopathology of BuHV-1 or BoHV-1 infection in Bubalus bubalis, we collected blood through iugulation and TG after decapitation. Samples from each TG were placed in a single 50-ml conical tube, and the tube were placed in a dry ice ethanol bath. TG samples were then stored at -80°C. Animals submitted for sampling were chosen within the > 5 year age category because considered at higher risk of infection. The presence of antibodies against BuHV-1 or BoHV-1 was investigated in 15 blood samples by enzyme-linked immunosorbent assay (anti-gB/gE blocking ELISA) (Idexx). Through the combined use of gB-gE ELISA tests, we assigned a specific infection status, for the BuHV1 infection status (gB-pos/gE-neg), and for the BoHV1 status (gB-pos/gE-pos) as reported by Scicluna et al. (2007). The ELISA results showed that 40% of tested samples was positive at BoHV-1 whereas 33.3% resulted to be BuHV-1 positive. Moreover 26.7% were BoHV-1 and BuHV-1 negative (gB-neg/gE-neg). Furthermore, we examined trigeminal ganglia tissues of the 15 animals. Oxidative status was assessed using the Reactive Oxygen Metabolites-derived compounds (d-ROMs) test and the antioxidant activity (anti- ROMs) test through spectrophotometric procedures. TG of seropositive or seronegative buffaloes were also used to measure the concentration of lipid peroxides using the Lipotiss test. We detected in TG of seropositive BuHV-1 animals a significantly reduction of d- ROMs values (P < 0.001) as well as anti-ROMs values (P < 0.001) compared with those in TG of seronegative animals. Whereas TG of seropositive BoHV-1 animals had significantly higher d-ROMs values (P < 0.001) and lower anti-ROMs values (P < 0.001) compared with those in TG of seronegative animals. Furthermore, the results of lipotiss test showed that samples of both seropositive BuHV-1 and BoHV-1 animals were significantly lower (P < 0.001; P < 0.01) compared with those of seronegative animals. Taken together, our preliminary results suggest that oxidative stress pattern and oxidative defence barrier are altered in latently infected TG compared with control uninfected TG. In particular, our results suggest that the presence of BuHV-1, virus species-specific, seems to induce a worsening balance in ROMs levels. Future studies are needed in order to assess the prognostic role of oxidative stress in trigeminal ganglia of Bubalus bubalis.

EVALUATION OF REACTIVE OXYGEN METABOLITES IN TRIGEMINAL GANGLIA OF ALPHAHERPESVIRUS SEROPOSITIVE ITALIAN MEDITERRANEAN BUFFALOES (BUBALUS BUBALIS)

Marullo, Annarosaria;TAFURI, SIMONA;CIANI, FRANCESCA;DELLA MORTE, ROSSELLA;F. P. Nocera;MALLARDO, KARINA;FIORITO, FILOMENA;DE MARTINO, LUISA
2014

Abstract

In nature, herpesviruses are mostly associated with a single host species. Italian Mediterranean buffaloes (Bubalus bubalis) is associated with Bubaline herpesvirus 1 (BuHV-1) but also with Bovine herpesvirus 1 (BoHV-1). Like different alphaherpesvirinae subfamily members, the primary site for BuHV-1 or BoHV-1 latency is sensory neurons within trigeminal ganglia (TG). Recently, it has been shown that cattle infected by BoHV-1 seemed to have more oxidative stress and low antioxidant defense (Durgut et al., 2013). Thus, to better understand the physiopathology of BuHV-1 or BoHV-1 infection in Bubalus bubalis, we collected blood through iugulation and TG after decapitation. Samples from each TG were placed in a single 50-ml conical tube, and the tube were placed in a dry ice ethanol bath. TG samples were then stored at -80°C. Animals submitted for sampling were chosen within the > 5 year age category because considered at higher risk of infection. The presence of antibodies against BuHV-1 or BoHV-1 was investigated in 15 blood samples by enzyme-linked immunosorbent assay (anti-gB/gE blocking ELISA) (Idexx). Through the combined use of gB-gE ELISA tests, we assigned a specific infection status, for the BuHV1 infection status (gB-pos/gE-neg), and for the BoHV1 status (gB-pos/gE-pos) as reported by Scicluna et al. (2007). The ELISA results showed that 40% of tested samples was positive at BoHV-1 whereas 33.3% resulted to be BuHV-1 positive. Moreover 26.7% were BoHV-1 and BuHV-1 negative (gB-neg/gE-neg). Furthermore, we examined trigeminal ganglia tissues of the 15 animals. Oxidative status was assessed using the Reactive Oxygen Metabolites-derived compounds (d-ROMs) test and the antioxidant activity (anti- ROMs) test through spectrophotometric procedures. TG of seropositive or seronegative buffaloes were also used to measure the concentration of lipid peroxides using the Lipotiss test. We detected in TG of seropositive BuHV-1 animals a significantly reduction of d- ROMs values (P < 0.001) as well as anti-ROMs values (P < 0.001) compared with those in TG of seronegative animals. Whereas TG of seropositive BoHV-1 animals had significantly higher d-ROMs values (P < 0.001) and lower anti-ROMs values (P < 0.001) compared with those in TG of seronegative animals. Furthermore, the results of lipotiss test showed that samples of both seropositive BuHV-1 and BoHV-1 animals were significantly lower (P < 0.001; P < 0.01) compared with those of seronegative animals. Taken together, our preliminary results suggest that oxidative stress pattern and oxidative defence barrier are altered in latently infected TG compared with control uninfected TG. In particular, our results suggest that the presence of BuHV-1, virus species-specific, seems to induce a worsening balance in ROMs levels. Future studies are needed in order to assess the prognostic role of oxidative stress in trigeminal ganglia of Bubalus bubalis.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11588/609463
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