Detection of indoleamine 2,3-dioxygenase 1-expressing cells in canine normal and tumor tissues

Cancer immunotherapy is a novel cancer treatment for canine tumors. Indoleamine 2,3-dioxygenase 1 (IDO1) is overexpressed in some human tumors and inhibits antitumor immunity. In this study, we comprehensively evaluated expression pattern of IDO1 and the nature of IDO1-expressing cells in canine normal and tumor tissues. In normal tissue samples, IDO1 expression was detected only in the lymph nodes, spleen, tonsil tissues, and colon tissues. In contrast, IDO1-positive tumor cells were observed in several tumor tissue types. This is the first study to evaluate IDO1 expression in canine normal and tumor tissues, and the results suggest that IDO1 is a promising target for novel cancer immunotherapy in dogs with tumors.     

Serological survey of antibodies to four pathogens in wild boars in Nagano Prefecture,Japan

Wild boars (Sus scrofa) captured or found dead in Nagano Prefecture were surveyed for antibodies to Aujeszky’s disease virus (ADV), Erysipelothrix rhusiopathiae, porcine reproductive and respiratory syndrome virus (PRRSV), and Toxoplasma gondii. While all 168 samples tested were negative for anti-ADV antibodies and all 140 samples tested were negative for anti-PRRSV antibodies, all 190 samples tested were positive for anti-E. rhusiopathiae antibodies and 12 of 180 samples were positive for anti-toxoplasma antibodies. These results suggest that since E. rhusiopathiae and T. gondii cause zoonotic diseases, in addition to wild boars being a potential source of infection for domestic pigs, caution should be taken when handling wild boars or eating wild boar meat because of the possibility of human infection.     

The effect of frequent milk feeding on abomasal curd formation of Holstein calves

In order to understand the effects of the automatic milk replacer feeding system on calf health, we examined the effect of frequent milk feeding on curd formation in the abomasum using ultrasonographic imaging. Eight male Holstein calves were divided into a milk-replacer group and a fresh milk group. Calves were fed twice a day to 12 days after birth (Period A). From 13 days, calves were fed six times a day (Period B). The abomasal fluid was taken by paracentesis. In both periods A and B, the fresh milk group formed bigger curds faster than the milk replacer group. The curd score of the milk replacer group in period B at 2 h was significant lower than those of both groups in period A. The pH in the abomasum was lower in period B than in period A. We could not identify the location of the abomasum in the milk replacer group during period B at almost time points. Our experiment suggests the possibility that frequent feeding of milk replacer causes incomplete hydrolysis of κ-casein as well as curd formation, thus reducing the digestibility compared to the feeding of the milk replacer twice per day or frequent feeding of fresh milk.     

Fatty acid‐binding protein expression in the gastrointestinal tract of calves and cows

Fatty acid‐binding protein (FABP) has high affinity for long‐chain fatty acids and appears to participate in the metabolism and intracellular transport of lipids. Liver‐ and intestinal‐type FABP (L‐FABP and I‐FABP, respectively) are expressed in the small intestine. However, in the gastrointestinal tract of ruminants, expression and localization of FABPs are unknown. In this study, we investigated the expression of I‐FABP and L‐FABP in the gastrointestinal tract of cattle. I‐ and L‐FABP had higher messenger RNA (mRNA) and protein expression levels in the duodenum and jejunum relatively to other gastrointestinal regions in both calves and cows. Furthermore, L‐FABP mRNA and protein expression were high in the colon. Both these protein types were confirmed to be in the cytosol of jejunal epithelial cells, where they were found in the villi rather than in the crypts. We concluded that duodenal and jejunal FABPs might be involved in the metabolism of fatty acids mainly in epithelial cells in cattle.     

Effect of supplementation of soy sauce oil and Ca salts of fatty acids on rumen fermentation,milk production and conjugated linoleic acid in milk of dairy cows

Eight cows were used to evaluate the effects of supplementation of soy sauce oil (SO) or Ca salts of fatty acids (FA) on rumen fermentation and milk production. The control diet (CO) consisted mainly of hay, corn silage and a concentrate. In the experimental diets, 400 g/day per cow of SO or FA (soybean oil and rapeseed oil) was supplemented to the CO diet. Experimental period for the three treatments was 14 days, and milk samples were taken during the last 2 days and rumen sample was taken on the last day. Dry matter intake was not affected by the treatments. The number of rumen protozoa at 0 h increased by SO and FA diets. Total volatile fatty acids at 2 h after feeding of SO diet was decreased compared to CO. The milk composition yield did not differ among treatments, although the percentages of fat and protein were decreased by SO and FA diets. The proportions of C8–C16 fatty acids in milk fat decreased, and those of C18 increased by SO and FA diets. The proportion of cis‐9, trans‐11 conjugated linoleic acid in milk fat by SO and FA diets increased by 120% and 135%, respectively. In spite of the slight suppression of rumen fermentation by SO diet, negative effects on feed intake and milk production were not detected.     

Equine nonneoplastic abnormal ovary in a draft mare with high serum anti-Müllerian hormone: a case study

We performed a standing hand-assisted laparoscopic ovariectomy in a draft mare that presented with high serum anti-Müllerian hormone (AMH) level and had an enlarged single cystic ovary. Histopathological examination revealed no tumor cell proliferation in the ovary, but the presence of a large ovarian cyst was confirmed. In the diagnosis of abnormal ovaries in mares, a comprehensive assessment should be performed, including the monitoring of ovarian morphology and biomarkers over time, to determine the disease prognosis and treatment plan. The case of this mare with a nonneoplastic abnormal ovary and increased serum AMH level was rare. We suggest that standing hand-assisted laparoscopic ovariectomy is useful for the removal of large ovaries in draft mares.     

The enhancement of Th1 immune response by anti-PD-L1 antibody in cattle infected with Mycobacterium avium subsp. paratuberculosis

Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic enteritis of ruminants. Previous studies have shown that programmed death-ligand 1 (PD-L1) is associated with the disease progression, and PD-L1 blockade activates MAP-specific Th1 responses in vitro. Here, we performed anti-PD-L1 antibody administration using 2 MAP-infected cattle at the late subclinical stage of infection. After administration, bacterial shedding was reduced or maintained at a low level. Additionally, MAP-specific Th1 cytokine production was upregulated, and CD69 expression was increased in T cells. Collectively, the treatment has a potential as a novel control method against Johne’s disease.     

Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies

A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies directed against a synthetic peptide with an amino-acid sequence related to the C-terminus of bovine myoglobin and the whole molecule of sodium dodecyl sulphate (SDS)-denatured bovine myoglobin was adapted for detecting bovine myoglobin in contaminated feeds. The ELISA employed bovine meat extract of a known myoglobin concentration as a calibration standard and had an limit of detection (LOD) of 3.54 ng/ml and an limit of quantification (LOQ) of 11.0 ng/ml corresponding to 0.022% and 0.067% (wt/wt) bovine meat-and-bone-meal (MBM) mixed in 20-fold-diluted feed extracts, respectively. A cut-off threshold of 20.6 ng/ml bovine myoglobin was set to simplify ELISA and facilitate quick assessment of test results without a tedious calibration process. The ELISA was able to detect bovine MBM in artificially prepared model feeds, mixed botanical feeds, mixed botanical feeds with skimmed milk, fish meal, pork meal and pork/chicken meal at 0.1% (wt/wt). It was also able to detect sheep MBM in test feeds, but showed no reactivity to swine MBM, chicken MBM, skimmed milk or gelatine of bovine origin. The advantages of this method are the quick and easy extraction protocol of proteins from test feeds, using 100 mM sodium sulphide and 0.6% sodium dodecyl sulphate in the extraction solution and the effective detection of bovine and sheep MBM at 0.1% (wt/wt).