Morphology and volume of Meibomian glands ex vivo pre and post partial tarsal plate excision,cryotherapy and laser therapy in the dog using microCT

Objective

To determine the morphology and volume of Meibomian glands (MG) of dogs with microCT before and after partial tarsal plate excision (PTPE), cryotherapy, and laser therapy.

Procedure

MicroCT scans were made of 12 upper lids (ULs) and lower lids (LLs) of 12 dogs. After undergoing PTPE, 10 ULs and LLs were scanned again, and one UL and one LL was scanned after laser therapy and one UL and one LL after cryotherapy.

Results

The length of the area containing MGs did not change pre- and post-PTPE, and cryo- or laser therapy. The mean number of MGs in the ULs and LLs was 30.50 and 29.42, respectively, and did not change during the procedures. The average length of one individual MG was 2.60 mm. The mean volume of MGs in the 10 ULs and LLs pre-PTPE was 21.45 and 17.2 mm³, respectively, and 12.84 and 11.25 mm³ in the UL and LL after PTPE, respectively. The mean volume of MGs decreased from 29.78 mm³ precryotherapy to 28.91 mm³ post-treatment and in the lower eyelid from 22.87 to 22.4 mm³ after cryotherapy. The mean volume of MGs in the UL and LL before laser therapy was 8.95 and 6.78 mm³, respectively, and after 9.25 and 6.38 mm³, respectively.

Conclusion

MicroCT is a valuable tool to determine the morphology and the volume of MGs and to demonstrate changes that occur after PTPE, laser-, and cryotherapy. There is no need for additional preparation, such as staining, of the specimen prior to scanning.     

兔出血症病毒在细胞培养和组织中的形态发生

在电镜下系统地观察了感染后的细胞培养和组织中兔出血症病毒（ Ｒ Ｈ Ｄ Ｖ）的形态发生。感染早期,在细胞核内可见电子致密颗粒（１５ ｎｍ ）和未成熟的病毒颗粒 （２５ ｎｍ ）。中期,在细胞核和胞浆内出现大量成熟的病毒颗粒（３４ ｎｍ ）,并发现部分核内病毒通过扩大的核膜孔、核膜溶解扩大的核孔和乳头状突起的核膜向胞浆释放。感染末期,核染色质消失,核内大量感染病毒清淅可见。最终细胞溶解,病毒颗粒释放至细胞间隙。提示 Ｒ Ｈ Ｄ Ｖ 是在核内复制和装配的,应归属于细小病毒科。本试验结果不排除同时存在另一种小 Ｒ Ｎ Ａ 病毒。     

Ultrasonographic anatomy of the atlanto‐occipital region and ultrasound‐guided cerebrospinal fluid collection in rabbits (Oryctolagus cuniculus)

Cerebrospinal fluid analyses are important for diagnosis of neurologic problems in rabbits and for translational research projects using rabbits as models. Blind puncture of the cisterna magna is the current standard technique for sampling cerebrospinal fluid in this species. However, the complexity and small size of the cisterna magna and surrounding structures are limitations of this technique. Aims of this prospective, anatomic, pilot study were to (1) describe the normal anatomy of the atlanto‐occipital region, (2) describe ultrasonographic anatomic landmarks, and (3) develop and evaluate a technique for ultrasound‐guided puncture of the cisterna magna for cerebrospinal fluid sampling in rabbits. Thirty healthy rabbits were included and the study was conducted in three stages. Three rabbit cadavers were used for the first stage of the study. Then, the second stage was completed using 13 rabbit cadavers. Finally, the third stage was completed in 14 live rabbits. The ultrasound‐guided puncture performed in 13 cadavers was successful at the first attempt in 10 cases, and at the second attempt in the remaining three cases. In the in vivo study, the ultrasound‐guided puncture was successful in all 14 cases, without signs of complications. Findings supported the use of ultrasound‐guided puncture of the cisterna magna as a safe technique that may be used routinely or when the sample of cerebrospinal fluid cannot be obtained with the blind technique in rabbits.     

Effects of limit‐feeding diets with different forage‐to‐concentrate ratios on nutrient intake,rumination, ruminal fermentation,digestibility, blood parameters and growth in Holstein heifers

The objective of this study was using a wide range of dietary concentrate levels to investigate the major effects of limit‐feeding on heifers. Twenty‐four Holstein heifers were blocked into six groups and fed with one of four diets containing different levels of concentrate (20%, 40%, 60% and 80% on a dry matter (DM) basis) but with same intakes of metabolizable energy for 28 days. Increasing levels of dietary concentrate caused decreased (P ≤ 0.02) intakes of dry matter (DMI) and neutral detergent fiber and total rumination time, but increased (P < 0.01) nonfiberous carbohydrates intake, ruminal concentrations of NH₃‐N, propionate and butyrate, and digestibility of DM and crude protein. Dietary concentrate levels had no significant effect on most plasma concentrations and body measurements. The corrected average daily gain (CADG) and feed efficency (ADG/DMI, CFE) were linearly increased (P < 0.01) with increasing dietary concentrate levels when gut fill impact was removed. In conclusion, heifers limit‐fed high concentrate diets increased most ruminal fermentation parameters, CADG and CFE with similar body growth and blood metabolites as heifers fed low concentrate diets, and had the potential to be used as an effective feeding strategy in dairy heifers.     

Effects of concentrate‐to‐forage ratios and 2‐methylbutyrate supplementation on ruminal fermentation,bacteria abundance and urinary excretion of purine derivatives in Chinese Simmental steers

This study evaluated the effects of dietary concentrate levels and 2‐methylbutyrate (2MB ) supplementation on performance, ruminal fermentation, bacteria abundance, microbial enzyme activity and urinary excretion of purine derivatives (PD ) in steers. Eight ruminally cannulated Simmental steers (12 months of age; 389 ± 3.7 kg of body weight) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement. Moderate‐concentrate (400 g/kg diet [MC ]) or high‐concentrate (600 g/kg diet [HC ]) diets were fed with or without 2MB (0 g/day [2MB ?] or 15.0 g/day [2MB +]). Dry matter intake and average daily gain increased, but feed conversion ratio decreased with the HC diet or 2MB supplementation. Ruminal pH decreased, but total volatile fatty acid increased with the HC diet or 2MB supplementation. Molar proportion of acetate and acetate‐to‐propionate ratio decreased with the HC diet, but increased with 2MB supplementation. Propionate molar proportion and ruminal NH ₃‐N content increased with the HC diet, but decreased with 2MB supplementation. Neutral detergent fibre degradability decreased with the HC diet, but increased with 2MB supplementation. Crude protein degradability increased with the HC diet or 2MB supplementation. Abundance of Ruminococcus albus , Ruminococcus flavefaciens , Fibrobacter succinogenes and Bufyrivibrio fibrisolvens as well as activities of carboxymethyl cellulase, cellobiase, xylanase and pectinase decreased with the HC diet, but increased with 2MB supplementation. However, abundance of Prevotella ruminicola and Ruminobacter amylophilus as well as activities of α‐amylase and protease increased with the HC diet or 2MB supplementation. Total PD excretion also increased with the HC diet or 2MB supplementation. The results suggested that growth performance, ruminal fermentation, CP degradability and total PD excretion increased with increasing dietary concentrate level from 40% to 60% or 2MB supplementation. The observed diet × 2MB interaction indicated that supplementation of 2MB was more efficacious for improving growth performance, ruminal fermentation and total PD excretion with promoted ruminal bacteria abundance and enzyme activity in the MC diet than in the HC diet.     

A novel surgical technique for enucleation in rabbits to reduce the risk of intra‐ and post‐operative orbital hemorrhage

A 10‐year‐old male castrated Holland Lop rabbit (Oryctolagus cuniculus) was presented for severe ulcerative stromal keratitis of the right eye and a luxated hypermature cataract and glaucoma of the left eye. Staged bilateral enucleation was elected. A LigaSure? electrosurgical bipolar vessel‐sealing device was used as a means to minimize intraoperative and post‐operative hemorrhage, especially that associated with the orbital venous plexus. The LigaSure? was used to ligate and transect all extraocular muscles, the optic nerve bundle, and the base of the third eyelid with no complications encountered. Overall, the LigaSure? was easy to use, resulted in minimal hemorrhage, and reduced surgery time. This is the first report of the use of a LigaSure? to aid in the enucleation of a rabbit. Although only positive results were achieved as an alternative to conventional methodologies, its use in clinical practice should be that of caution until a larger study evaluating the long‐term results is performed.     

Ultrastructure of the tubular glands in the isthmus region of the oviduct in laying and natural moulting commercial egg‐type chickens

The study investigated the ultrastructural characteristics of tubular gland and duct cells, as well as luminal gland cells in the isthmus region of the oviduct of laying and natural moulting hens. Tubular glands in laying birds were composed of type 1 and 2 cells. Based on the preponderance of each cell type, in relation to the location of a developing egg in the oviduct of the domestic fowl, these gland cells may represent different functional states of the same cell. The findings of the study on natural moulting birds suggest that autophagy is a process confined to the early stages of degeneration, while necrosis occurs in the terminal stages.     

Simple method for isolation of glyceraldehyde 3‐phosphate dehydrogenase and the improvement of myofibril gel properties

Porcine glycoliytic enzyme, glyceraldehyde 3‐phosphate dehydrogenase (G3PD) was prepared effectively by a combination of ethylene diamine tetra‐acetate (EDTA) pretreatment and affinity purification. After salting out of porcine sarcoplasmic proteins (SP) with ammonium sulfate at 75% saturation, the obtained supernatant (SP‐f3) was treated with EDTA, leaving G3PD in the supernatant (G3PD‐E) and most other SPs in the precipitate. At that time, the separation of G3PD‐E required more than 20 mmol/L EDTA. G3PD‐E was then subjected to affinity purification by batchwise method using blue‐sepharose CL‐6B, and purified G3PD (G3PD‐AP) was obtained using 2 mol/L potassium chloride (KCl) as an eluent. Texture analysis showed that the hardness, adhesiveness and gumminess of the myofibril gel at 0.2‐mol/L NaCl increased with the addition of G3PD‐AP. Scanning electron microscopy revealed that the G3PD‐AP reinforced the gel network of the myofibril. However, scanning electron micrograph analysis showed that the network‐structure of the gel by the addition of G3PD‐AP developed in a different manner from that by adding 0.6 mol/L NaCl. These results showed that glycolytic enzyme, G3PD, contributes to the improvement of the rheological properties of meat products.     

Branched‐chain amino acid ratios in low‐protein diets regulate the free amino acid profile and the expression of hepatic fatty acid metabolism‐related genes in growing pigs

Liver metabolism is affected by nutrients. The aim of this study was to explore the effects of low‐protein diets (17% crude protein, CP) supplemented with branched‐chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile) and valine (Val), on hepatic amino acid profile and lipid metabolism in growing pigs. The ratio of Leu : Ile : Val in all groups was 1 : 0.51 : 0.63 (20% crude protein, CP), 1 : 1 : 1 (17% CP), 1 : 0.75 : 0.75 (17% CP), 1 : 0.51 : 0.63 (17% CP) and 1 : 0.25 : 0.25 (17% CP) respectively. Results revealed that compared to the positive control group (1 : 0.51 : 0.63, 20% CP), the low‐protein diets significantly augmented the concentrations of most essential amino acids and non‐essential amino acids (p < .05), with the greatest values observed in the 1 : 0.25 : 0.25 group. Moreover, relative to the control, the low‐protein diets with the Leu : Ile : Val ratio ranging from 1 : 0.75 : 0.75 to 1 : 0.25 : 0.25 markedly downregulated the mRNA abundance of acetyl‐CoA carboxylase (ACC), lipoprotein lipase (LPL) and fatty acid‐binding protein 4 (FABP‐4) (p < .05), and upregulated the mRNA expression of hormone‐sensitive lipase (HSL), peroxisome proliferator‐activated receptor‐g coactivator‐1α (PGC‐1α), uncoupling protein 3 (UCP3) and liver carnitine palmitoyltransferase 1 (L‐CPT‐1) (p < .05). Therefore, our data suggest that protein‐restricted diets supplemented with optimal BCAA ratio, that is, 1 : 0.75 : 0.75–1 : 0.25 : 0.25, induce a shift from fatty acid synthesis to fatty acid oxidation in the liver of growing pigs. These effects may be associated with increased mitochondrial biogenesis.     

Genetic analysis of fat‐to‐protein ratio,milk yield and somatic cell score of Holstein cows in Japan in the first three lactations by using a random regression model

We estimated the genetic parameters of fat‐to‐protein ratio (FPR) and the genetic correlations between FPR and milk yield or somatic cell score in the first three lactations in dairy cows. Data included 3 079 517 test‐day records of 201 138 Holstein cows in Japan from 2006 to 2011. Genetic parameters were estimated with a multiple‐trait random regression model in which the records within and between parities were treated as separate traits. The phenotypic values of FPR increased soon after parturition and peaked at 10 to 20 days in milk, then decreased slowly in mid‐ and late lactation. Heritability estimates for FPR yielded moderate values. Genetic correlations of FPR among parities were low in early lactation. Genetic correlations between FPR and milk yield were positive and low in early lactation, but only in the first lactation. Genetic correlations between FPR and somatic cell score were positive in early lactation and decreased to become negative in mid‐ to late lactation. By using these results for genetic evaluation it should be possible to improve energy balance in dairy cows.     

Effects of yeast culture supplementation and the ratio of non‐structural carbohydrate to fat on growth performance,carcass traits and the fatty acid profile of the longissimus dorsi muscle in lambs

The effects of yeast culture (YC) supplementation and the dietary ratio of non‐structural carbohydrate to fat (NSCFR) on growth performance, carcass traits and fatty acid profile of the longissimus dorsi (LD) muscle in lambs were determined in a 2 × 3 full factorial experiment. Thirty‐six Small‐tailed Han lambs were randomly divided into six groups with six replicates per group. The lambs were fed one of the six pelleted total mixed rations (TMRs) for 60 days after 15 adaption days. The six rations were formed by two NSCFRs (11.37 and 4.57) and three YC supplementation levels (0, 0.8 and 2.3 g/kg dietary dry matter). The average daily gain (ADG), dry matter intake (DMI) and feed conversion ratio (FCR) data of each lamb were recorded and calculated. All the lambs were slaughtered for determining carcass traits and fatty acid profile of the LD muscle. DMI was significantly increased (p < 0.05) in a quadratic fashion with 0.8 g/kg of YC supplementation. Carcass weight (CW) and dressing percentage (DP) were significantly increased (p < 0.05) in a linear fashion with 2.3 g/kg of YC supplementation. Animals fed with high‐NSCFR diet had higher (p < 0.05) contents of myristoleic acid (C14:1), pentadecanoic acid (C15:0) and cis‐10‐heptadecenoic acid (C17:1), and lower (p < 0.05) stearic acid (C18:0) content in LD muscle than those fed with low‐NSCFR diet. Moreover, ADG, growth rate (GR), backfat thickness (BFT), percentages of crude fat (CF) and crude protein (CP), SFAs, MUFAs and PUFAs in LD muscle, were significantly affected (p < 0.05) by interaction of dietary NSCFR and supplemental YC level. Overall, YC not only improved the growth performance and carcass traits of the animals but also modified the fatty acid profile of the LD muscle. Furthermore, the effects of YC supplementation may depend on dietary compositions.