Expression and single nucleotide polymorphisms of vitamin D receptor gene in cows with puerperal metritis in Egypt.

The aim of this work is to study the expression profile of the vitamin D receptor (VDR), 1-α hydroxylase enzyme, and chemokine regulated on activation normal T-cell expressed and secreted genes (RANTES) genes in dairy cows with puerperal metritis, as well as to study the association between polymorphisms in the VDR gene and occurrence of such disease condition, which is considered a key to advances in the preventive medicine for such a problem in the future. Blood samples were collected from 60 dairy cows; from which 48 dairy cows proved to suffer puerperal metritis and other 12 apparently healthy recent parturient dairy cows were selected randomly for assessment the fold change variation in the expression profiles of the studied genes. Depending on the clinical examination, the normal cut-off point of the serum calcium, phosphorus, glucose, magnesium, potassium, sodium, and chloride, and the normal reference values of parathyroid hormone (PTH), 18 out of 48 dairy cows suffered puerperal metritis with a normal metabolic profile, which underwent further molecular analysis. In the studied dairy cows that suffered puerperal metritis with a normal metabolic profile, the expression pattern of the VDR and RANTES genes showed a significant up-regulation (P < 0.05), whereas the expression pattern of 1α-hydroxylase enzyme gene showed a significant down-regulation (P < 0.05) in comparison with those of healthy recent parturient dairy cows. The sequenced regions of the VDR gene (exon 7, partial intron 6, partial intron 7) didn't exhibit polymorphisms in their nucleotide sequence, and consecutively their amino acid sequence when compared to the published sequence on the gene bank. The results propose that the VDR gene (exon 7, partial intron 6, partial intron 7) polymorphisms do not play a role in the pathogenesis of puerperal metritis in dairy cows, excluding a role of the vitamin D in puerperal metritis.

Utilizing attached hard ticks as pointers to the risk of infection by Babesia and Theileria species in sika deer (Cervus nippon yesoensis), in Japan.

Ticks are hematophagous ectoparasites that have a significant impact on their animal hosts. Along with mosquitoes, they are the main arthropod vectors of disease agents in domestic animals, wildlife and humans. To investigate the occurrence and prevalence of piroplasmids in ticks, DNA was extracted from 519 hard ticks collected from 116 hunted Hokkaido sika deer (Cervus nippon yesoensis). The success of the DNA extraction was confirmed by touchdown PCR targeting the mitochondrial 16S rDNA gene of ticks. Touchdown PCR and reverse line blot (RLB) hybridization targeting the 18S rRNA gene were used to detect 14 piroplasm species. All hard ticks parasitizing Hokkaido sika deer were identified as belonging to the genera Ixodes and Haemaphysalis. In total 163 samples (31.4%) were positive for Babesia and Theileria spp. among tick species according to RLB hybridization. Tick DNA hybridized to the oligonucleotide probes of Theileria sp. Thrivae (27.0% of ticks; 140/519), Theileria capreoli (10.6%; 55/519), Babesia divergens-like (1.7%; 9/519), Babesia sp. (Bab-SD) (0.6%; 3/519), Babesia microti U.S. (0.4%; 2/519), and B. microti Hobetsu (0.4%; 2/519). The partial sequencing and phylogenetic analyses of the 18S rRNA gene confirmed the RLB hybridization results. Further investigations are needed to reveal the epidemiology and respective vectors of these pathogens.

Molecular identification and characterization of piroplasm species in Hokkaido sika deer (Cervus nippon yesoensis), Japan.

Babesia and Theileria species are tick-borne protozoan parasites that have a veterinary and zoonotic importance. In order to investigate the prevalence and genetic diversity of these parasites, a total of 269 sika deer blood DNA samples collected from Hokkaido, Japan, were examined for Babesia and Theileria species by touch-down PCR targeting the 18S rRNA gene. Reverse line blot (RLB) hybridization was then used to detect 12 piroplasm species. The results revealed that 95.5% (257/269), 94.1% (253/269), 14.1% (38/269), 87.7% (236/269) and 11.5% (31/269) of the examined PCR products hybridized with the probes which were designed to detect all Babesia and Theileria spp., all Theileria spp., all Babesia spp., Theileria sp. Thrivae and Babesia divergens-like, respectively. The 18S rRNA gene partial sequences were divided into Theileria sp. Thrivae, T. capreoli, B. divergens-like and an undescribed Babesia species. This study showed the first detection of the undescribed Babesia sp. from Japan. Therefore, more studies are required to understand the ecology of the newly detected tick-borne pathogens in Hokkaido.

Effect of selenium and vitamin C on clinical outcomes, trace element status, and antioxidant enzyme activity in horses with acute and chronic lower airway disease. A randomized clinical trial.

Excess production of reactive oxygen species is involved in the pathogenesis of airway disorders in horses. Trace element antioxidants have a beneficial role in oxidant/antioxidant balance. The aim of the present study was to evaluate the effect of a combination of sodium selenite and ascorbic acid on clinical outcome, antioxidant enzymes, and trace elements status in horses with lower airway disease. For this purpose, 40 draft horses with lower airway disease were randomly selected (acute, n = 20; chronic, n = 20). Both acute and chronic cases were randomly allocated into two subgroups (ten each). Groups 1 and 2 were the horses with acute disease, while groups 3 and 4 were chronically ill. For all groups, each horse was administered antibiotic, non-steroidal anti-inflammatory, and mucolytic drug. In addition, groups 2 and 4 were injected with 15 mg/kg sodium selenite and 30 mg/kg ascorbic acid every 24 h for successive 4 weeks. Venous blood samples were obtained from diseased horses on three occasions; at first examination, and at 2 and 4 weeks post-treatment. Clinically, antioxidant supplementation improved the clinical signs with significant decrease (p < 0.05) of the clinical index score in both acute and chronic cases. In supplemented groups compared with non-supplemented, there was a significant increase (p < 0.05) in the levels of copper, zinc, selenium, and iron as well as in the activity of glutathione-S-transferase and catalase. Meanwhile, there was a significant decrease (p < 0.05) in the levels of manganese, malondialdehyde, hydrogen peroxide, and low-density lipoprotein and in the activity of glutathione reductase. The results of the present study indicate that administration of sodium selenite and ascorbic acid may have beneficial effect on clinical outcome and antioxidant balance in horses with acute and chronic lower airway disease.

Antioxidant trace elements in serum of draft horses with acute and chronic lower airway disease.

The aim of the present study was to evaluate the oxidative stress level and antioxidant trace elements status associated with lower airway disease in draft horses. For this purpose, venous blood samples were obtained from draft horses exhibiting signs of lower respiratory tract disorders (n = 83) and from control group (n = 20). Serum trace elements including selenium (Se), copper (Cu), zinc (Zn), manganese (Mn), and iron (Fe) were assayed. Serum malondialdehyde (MDA) and low-density lipoprotein (LDL) levels as well as plasma hydrogen peroxides (H2O2) concentration and activity of plasma glutathione reductase (GR), glutathione-S-transferase (GST) and catalase (CAT) were measured. There was a significant (p < 0.05) decrease of Se, Cu, Zn, and Fe in diseased horses compared with healthy ones, but the Cu/Zn ratio and Mn were increased (p < 0.05). Se was significantly (p < 0.05) decreased in chronically affected horses compared with acute cases, but Mn was increased (p < 0.05). There was an increase of MDA, LDL, and H2O2 levels and GR activity in diseased cases compared with healthy horses. However, there was a significant (p < 0.05) decrease of GST and CAT activity. MDA and LDL levels were increased (p < 0.05) in horses with chronic respiratory disease compared to acute cases, but CAT activity was decreased (p < 0.05). In horses with acute lower airway disease, there was a negative correlation between GR and H2O2 (r = -0.458), and LDL and CAT (r = -0.816). However, in chronic disease, a negative correlation was recorded between Se and MDA (r = -0.590). The results of the present study indicate that oxidative stress, with alteration of antioxidant trace element levels, is a feature of respiratory disease in draft horses.

Ketosis in buffalo (Bubalus bubalis): clinical findings and the associated oxidative stress level.

As little is known about the oxidant/antioxidant status in buffalo with ketosis, the present study was delineated to assess the oxidative stress level associated with clinical ketosis in water buffalo. A total of 91 parturient buffalo at smallholder farms were studied (61 suspected to be ketotic and 30 healthy). Clinical and biochemical investigations were carried out for each buffalo. Based on clinical findings and the level of beta-hydroxybutyrate (BHB), buffalo were allocated into ketotic (42), subclinical cases (19). Clinically, there was an association between clinical ketosis and anorexia (p<0.001), constipation (p<0.001), decreased milk yield (p<0.001), ruminal stasis (p<0.001), and loss of body condition (p<0.01). Biochemically, in clinical ketosis compared with subclinical and control cases, there was a significant increase (p<0.05) of BHB, malondialdehyde (MDA), nitric oxide (NO), aspartate aminotransferase (AST), L-alanine aminotransferase (ALT). However, there was a significant decrease of glucose, phosphorus, magnesium,total cholesterol and HDL-cholesterol. There was a positive correlation between BHB and MDA (r=0.433), BHB and NO (r=0.37), MDA and NO (r=0.515), and Glucose and phosphorus(r=0.521). However, there was a negative correlation between BHB and glucose (r= -0.341) and HDL and NO (r= -0.379). The result of the present study indicates that hyperketonemia in buffalo is associated with an increase of oxidative stress levels. Further studies need to be done on the efficacy of antioxidants as an ancillary treatment to relief the oxidative stress caused by ketosis.