Use of comparative proteomics to identify the effects of creatine pyruvate on lipid and protein metabolism in broiler chickens

Four hundred male chickens were selected to study the effects of pyruvate (Pyr), creatine pyruvate (CrPyr) and creatine (Cr) on the expression of hepatic mitochondrial and cytoplasm proteins associated with lipid and protein metabolism. Mitochondrial purification was accomplished using the two-step differential centrifugation and density gradient method, and the activities of organelle-specific marker enzymes were determined to assess the purity of the mitochondria. Proteins were extracted and fractionated by two-dimensional electrophoresis and the differential protein spots were assessed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. CrPyr reduced fatty acid accumulation by down-regulating adipose differentiation-related protein, inhibited ATP synthase expression, and reduced cholesteryl ester transfer protein (CETP) expression, thus reducing the levels of high density lipoprotein and triglycerol (TG) levels (thereby lowering fat and cholesterol deposition). CrPyr increased the expression of eukaryotic translation initiation factor (eIF) 2B, calreticulin (CRT) and eIF3a, thus promoting protein synthesis. CrPyr up-regulated the expression of fatty acid-binding proteins, CETP and apolipoprotein A-IV in cytoplasmic extracts, and these proteins accelerated the decomposition of fatty acids and TG, thus reducing fat deposition. In conclusion, CrPyr plays an important role in lipolysis and protein synthesis, and this effect was more pronounced than was the effect of Pyr and Cr.     

Septicaemia secondary to infection by Corynebacterium macginleyi in an Indian python (Python molurus)

A seven-year-old female Indian python (Python molurus) weighing about 35kg was euthanased after several clinical episodes of stomatitis, pneumonia, ophthalmitis and dystocia over a period of four years. The animal had been maintained in a terrarium in a circus truck at an adequate temperature. During shows, however, the snake was considered to be exposed to stressful conditions for several hours at a time at low temperatures and with noise and bright lights. A post-mortem examination indicated ulcerative stomatitis, osteomyelitis, severe pneumonia and numerous granulomata and multifocal necrosis in stomach and spleen. Corynebacterium macginleyi was isolated in pure culture from the ulcerative stomatitis, and mixed with Stenotrophomonas maltophilia from the lungs and spleen. The findings indicated that the snake had died from a septicaemic process caused by C. macginleyi, probably originating from the stomatitis. The role of S. maltophilia as a secondary agent is discussed. The stress of the circus show and poor husbandry may have predisposed the animal to infection and septicaemia. This is the first report of C. macginleyi causing disease in a snake.     

猪圆环病毒2型感染对猪繁殖与呼吸综合征灭活疫苗体液免疫的影响

本试验采用ELISA方法对单独接种猪繁殖与呼吸综合征病毒(PRRSV)变异株灭活苗组(HP组,n=3)和PCV2感染且出现病毒血症后接种PRRSV变异株灭活苗组(PCV2/HP组,n=3)不同时相血清中的PRRSV抗体进行检测;并对HP和PCV2/HP组血清中PCV2特异的抗体和核酸分别进行ELISA和荧光定量PCR检测。结果表明,在首免后70 d HP组血清平均抗体效价极显著高于PCV2/HP组(P<0.01);首免后56、63和77 d HP组平均抗体效价明显高于PCV2/HP组。其中在首免后56和63 d HP组抗体阳性率均达67%(2/3),PCV2/HP组在相应时相抗体阳性率为0;在首免后70和77 d HP组抗体阳性率均达100%(3/3),PCV2/HP组在相应时相抗体阳性率仅为0和33%(1/3)。结果提示PCV2感染可在一定程度上抑制PRRSV变异株(JXA1)特异性的抗体反应。     

Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.     

Effect of oxytocin infusion on luteal blood flow and progesterone secretion in dairy cattle

The objective of this study was to investigate the effects of oxytocin infusion on corpus luteum (CL) function during early to mid-diestrus by measuring luteal size (LS) and luteal blood flow (LBF) along with plasma levels of progesterone (P4) and prostaglandin metabolites (13,14-dihydro-15-keto-prostaglandin F_2α, PGFM). On day (D) 7 of the estrus cycle (D1 = ovulation), seven cows received 100 IU of oxytocin (OXY) or placebo (PL) following a Latin square design. LS and LBF increased in both groups over time and no differences were observed between the groups. PGFM did not differ either within the groups over time or between the groups at any time point. P4 of the OXY group was higher compared to that of the the PL group 360 min after the infusion (p = 0.01) and tended to be higher at the time points 450 min, 48 h, and 72 h (all p = 0.08). Results from this study support the hypothesis that OXY is not directly involved in the mechanism(s) governing blood flow of the CL and has no remarkable effects either on luteal size or P4 and PGFM plasma levels. Further investigation is needed to elucidate the role of OXY in CL blood flow during early and late luteal phases.     

In vitro antiviral activity of dehydroepiandrosterone and its synthetic derivatives against vesicular stomatitis virus

In this work the antiviral activity of 20 dehydroepiandrosterone (DHEA) analogs with different substituents at positions C-3, C-15, C-16 and C-17 were evaluated against vesicular stomatitis virus (VSV) in Vero cell cultures. The selectivity indexes (SI) obtained with DHEA and epiandrosterone (EA) were 50 and 72.6, respectively. The work showed that the compounds 21-norpregna-5,17(20)-dien-3β,16α-diyl-diacetate, 17,17-ethylendioxyandrostan-5,15-dien-3β-ol and 3β-hydroxypregn-17(20)-en-16-one had higher SI values than ribavirin, which was used as a reference drug. The antiviral mode of action of DHEA was also investigated against VSV replication in Vero cells, and time of addition experiments showed that DHEA mainly affected a late event in the virus growth cycle. Analysis of RNA and protein synthesis indicated that DHEA adversely affected positive strand RNA synthesis and viral mature particle formation.     

Diagnosis and treatment of presumptive pyelonephritis in an Asian elephant (Elephas maximus).

A 37-yr-old female Asian elephant (Elephas maximus) presented with anorexia, restlessness, and dark-colored urine. Urinalyses showed hematuria, leukocyturia, isosthenuria, proteinuria, granular casts, and no calcium oxalate crystals. Bloodwork revealed azotemia. Urine culture revealed a pure growth of Streptococcus zooepidemicus resistant to sulfamethoxazole-trimethoprim but susceptible to cephalosporins. A presumptive diagnosis of pyelonephritis was made based on bloodwork, urinalysis, and urine culture. The animal was treated with intravenous ceftiofur, and intravenous and per rectum fluids were given for hydration. The elephant's attitude and appetite returned to normal, the abnormal blood parameters resolved, and urinary calcium oxalate crystals reappeared after treatment, supporting presumptive diagnosis. Follow-up ultrasonography revealed an abnormal outline of both kidneys with parenchymal hyperechogenicity and multiple uterine leiomyomas.     

Quantification of pulmonary to systemic flow ratio by a Doppler echocardiographic method in the normal dog: Repeatability, reproducibility, and reference ranges

Objective

To determine if the pulmonary to systemic flow ratio (Qp/Qs) could be assessed in healthy awake dogs using Doppler echocardiography.

Background

Qp/Qs could provide reliable information in assessing the severity of intracardiac shunts (ICS) by quantifying pulmonary overcirculation. Qp/Qs has been validated against electromagnetic flowmeter methods in experimental canine models. However, its clinical applicability in awake dogs has never been assessed.

Animals, materials and methods

Six healthy dogs were used to determine the repeatability and reproducibility of the technique (Study 1); Qp/Qs was then prospectively assessed in 50 healthy dogs (Study 2). In both studies Qp/Qs was calculated in awake animals using a standardized Doppler echocardiographic method.

Results

Within- and between-day coefficients of variation for Qp/Qs were <10% (Study 1). For Study 2, a relatively wide range of Qp/Qs was found (reference range = 0.71-1.29; mean ± SD = 1.00 ± 0.15).

Conclusions

Qp/Qs can be assessed with good repeatability and reproducibility in healthy dogs. However, the wide range of Qp/Qs obtained in the healthy population may suggest a limited usefulness of this variable for accurately assessing ICS severity in diseased animals. This needs to be assessed in further prospective and longitudinal studies including a large number of animals with ICS of various grades.     

The Relationship Between the Positive Identification of the Embryo Proper in Equine Pregnancies Aged 18-28 Days and its Future Viability: A Field Study

Early embryonic loss (EEL) negatively affects the reproductive efficiency of equine reproduction. A sign of future EEL is when the embryo proper (EP) fails to develop within the embryonic vesicle after 30 days of gestation. The earlier the identification of impending EEL, the earlier the mare can be rebred to allow a second chance of pregnancy. The objectives of this study were to determine the percentage of embryonic vesicles with a visible EP at 18-28 days of gestation and to study the association between the presence/absence of the EP at different days of gestation and the future viability of the pregnancy. A total of 1,256 pregnancies were identified and followed by transrectal B-mode ultrasonography 12-45 days post ovulation in mares of the same Thoroughbred farm. The identification of the EP was attempted once during days 18-28. Pregnancy reconfirmation was performed on days 35-45. The percentage of vesicles with an EP increased gradually from days 18 (2.8%) to 21 (86.9%) (P < .05). From day 20 onward, the EEL rate of mares with vesicles without an EP was significantly higher (P < .05) than that of vesicles with a positive identification of an EP. In conclusion, the EP of the equine vesicle can be identified reliably with B-mode ultrasonography in the majority of mares (>71%) on day 20 of gestation. The lack of a positive identification of an EP from day 20 onward is associated with a higher EEL rate.     

Changes in nutrient balance,methane emissions,physiologic biomarkers,and production performance in goats fed different forage-to-concentrate ratios during lactation

The objective was to determine the effect forage-to-concentrate (F:C) ratio and stage of lactation on methane emissions, digestibility, nutrient balance, lactation performance, and metabolic responses in lactating goats. Twenty Murciano-Granadina dairy goats were used in an experiment divided into 3 periods: early (30 d), mid (100 d), and late (170 d) lactation. All goats were fed a diet with 35:65 F:C (FCL) during early-lactation. Then, 1 group (n = 10 goats) remained on FCL through mid- and late-lactation while the other group (n = 10 goats) was fed a diet with 50:50 F:C at mid-lactation (FCM) and 65:35 (FCH) at late lactation. A greater proportion of concentrate in the diet was associated with greater overall intake and digestibility (P < 0.05). Energy balance was negative in early-lactation (−77 kJ/kg of BW^0.75, on average) and positive for FCL at mid- and late-lactation (13 and 35 kJ/kg of BW^0.75, respectively). Goats fed FCM and FCH maintained negative energy balance throughout lactation. Plasma concentrations of non-esterified fatty acids at mid-lactation were greater for FCM than FCL (680 mEq/L), and at late-lactation concentrations were greater for FCH and FCL (856 mEq/L). A similar response was detected for plasma β-hydroxybutyrate. Methane emission was greater (P < 0.05) for FCM than FCH (1.7 g CH₄/d). This study demonstrated that differences in F:C across stages of lactation lead to distinct metabolic responses at the level of the rumen and tissues.