Evaluation of a wireless ambulatory capsule (SmartPill®) to measure gastrointestinal tract pH, luminal pressure and temperature, and transit time in ponies

Reasons for performing study: This study investigated the use of a wireless ambulatory capsule (WAC; SmartPill^® pH.p GI Monitoring System) to determine WAC‐gastric emptying time (GET) in ponies. Objectives: To measure WAC‐GET and compare it to those findings with GET assessed by nuclear scintigraphy (S‐GET). Hypothesis: WAC‐GET will be slower than S‐GET, but will be significantly correlated. Methods: Seven healthy adult mixed‐breed pony mares were used in this study. Feed was withheld for 12 h prior to the WAC administration. After administration, a complete‐feed diet was fed to allow the WAC to pass into the stomach. Luminal pH, temperature and pressure were collected by a modified receiver secured to the pony. Once the pH reached a value of ≥8.0, it was determined that gastric emptying had occurred, and ponies were fed grass hay. After 5 days, data were downloaded and analysed using proprietary software. During the second period of the study, after at least 2 weeks, 4 of the ponies underwent a standard S‐GET test. Results: The WAC was successfully administered, and data were collected from all ponies. The mean percentage of data packets collected by the receiver was 84.9 ± 3.51% (range 66.8–95.1%). Mean WAC‐GET was 7.38 h (range 0.15–46.65 h). Mean gastric pH was 4.75 (range 2.07–6.99). Mean small intestinal transit time was 4.6 h. The mean pH for the small intestine was 8.0. The mean S‐GET time (in hours) when 10% of the radioactive feed is present in the stomach (T‐90%) was 2.3 h. The S‐GET did not correlate significantly with the WAC‐GET. Conclusions and potential relevance: The WAC was safely administered to ponies, and data were collected using a modified receiver. The WAC‐GET varied considerably between ponies, but was ≤3 h in 5 of the 6 ponies. The WAC used in this study provided a noninvasive technique that produced novel information about the pony gastrointestinal tract, but owing to the substantial variability in GET values and long transit time it may not be a reliable clinical tool at this time.     

Effect of a soluble polysaccharide (carboxy methyl cellulose) on the physico‐chemical conditions in the gastrointestinal tract of broilers

1. The effects of an indigestible soluble polysaccharide (carboxy methyl cellulose: CMC) on broiler performance (body weight gain, food and water intake) and on chyme characteristics (moisture content, viscosity, pH, osmolality and retention time) in broilers were studied.

2. In semi‐synthetic diets 0, 5.0 and 10.0 g/kg of cellulose was replaced by CMC on weight basis. These diets were fed to male broilers from 3 to 5 weeks of age.

3. When 10.0 g/kg CMC was included in the diet, food intake and body weight gain were reduced, compared to the 0, and 5.0 g/kg CMC diets. Food:gain ratio and water intake were increased at each CMC concentration. The significant quadratic response showed an increased response per 5.0 g/kg dietary CMC at the higher CMC concentration.

4. The viscosity in the supernatant of the chyme was linearly increased in all intestinal segments by CMC. A quadratic increase was observed in the crop. In the lower ileum, differences between the 5.0 and 10.0 g/kg CMC diets were not significant.

5. CMC increased the mean retention time of chromium in the duodenum and in the upper jejunum, and reduced the maximal rate of marker excretion. The transit time (first appearance of the marker in the excreta), however, was significantly reduced at the highest CMC concentration.

6. The osmolality of the intestinal fluid decreased less as the chyme moved from the duodenum into the lower ileum, with increasing concentrations of CMC.

7. The ileal pH was reduced linearly by the CMC content of the diet.

8. Based on a higher moisture content of the chyme in the CMC‐fed birds, and the higher jejunal and ileal osmolarities in those birds, it was concluded that the efficiency of both digestion and absorption was reduced by CMC inclusion in broiler diets.     

Expression and Localization of Integrin αv in Different Parts of the Fallopian Tube in Reproduction Cycle of Yak

The objective of this study was to explore the expression differences of integrin αv in different parts of the fallopian tube at different reproduction stages of yak, and provide some important theoretical foundations for understanding the effect of integrin αv on the reproductive performance of yak. The samples of the tubal umbrella, ampulla, and isthmus of healthy female yaks (3-6 years old) during follicular, luteal and pregnancy stages were collected, the fallopian tubes of the follicular phase, the luteal phase and the pregnancy phase were divided into 9 groups according to the umbrella, ampulla and isthmus. Real-time PCR (qRT-PCR) and Western-blotting (WB) were used to detect the expression of integrin αv gene and protein. The expression of integrin αv protein was localized by immunohistochemistry. Results: 1) qRT-PCR results showed that, during the follicular phase, the expression of the integrin αv gene in the umbrella of the fallopian tube was the highest and extremely significantly higher than those in the ampulla and isthmus (P<0.01). During the luteal phase, the expression of the integrin αv gene in the isthmus of the fallopian tube was the highest and extremely significantly higher than those in the umbrella and ampulla (P<0.01). During the pregnancy phase, the expression of the integrin αv gene in the umbrella of the fallopian tube was the highest and extremely significantly higher than those in the ampulla and isthmus (P<0.01). 2) Western-blotting results showed that, during the follicular phase, the expression of integrin αv protein in the umbrella of the fallopian tube was the highest and extremely significantly higher than those in the ampulla and isthmus (P<0.01). During the luteal phase, the expression of integrin αv protein in the isthmus of the fallopian tube was the highest and extremely significantly higher than those in the umbrella and ampulla (P<0.01). During the pregnancy phase, the expression of integrin αv protein in tubal isthmus and umbrella was not significantly different, but these were extremely significantly higher than that in the ampulla (P<0.01). 3) The results of immunohistochemistry showed that integrin αv protein was positively expressed in ciliated cells, secretory cells, basal cells, muscular layers and serous glands of the fallopian tube umbrella, ampulla and isthmus. The results indicated that the expression of integrin αv in different parts of the fallopian tube at different reproduction stages of the yak were significantly different, which showed that αv might be involved in a series of reproductive processes such as fertilization and early embryonic development, the results provided basic information for the study of yak's reproductive performance.     

Quantitative effects of pelleting on performance,gastrointestinal tract and behaviour of meat‐type chickens

1. In an attempt to quantify the effects of “degree” of pelleting, two experiments were conducted. Diets were prepared by mixing together a mash composed mainly of maize (experiment 1) or sorghum (experiment 2) with soft pellets, or soft pellets mixed with hard pellets.

2. The pelleting degrees (PDs) were as follows: 0 mash; 0·5 mixture of soft pellets and mash 1 to 1; 1 soft pellets pelleted once; 1·5 mixture of soft and hard pellets 1 to 1; 2 hard pellets pelleted twice.

3. In experiment 2, the weight and length of the digestive organs were determined as well as digestive enzyme activities. In both experiments, the behaviour recorded was eating, standing, sitting and drinking.

4. Food intake and body weight gain were related to the degree of pelleting in a curvilinear manner. PD had a positive effect up to a peak (1 to 1·5 PD), after which its effect decreased. Food efficiency was not related to PD. In experiment 1, food efficiency of PDs 1 to 2 were superior to PDs 0 to 0·5 and in experiment 2, PDs 1·5 to 2 were superior to PD 0.

5. The relative weight of the gizzard was reduced by pelleting, whereas pelleting increased the relative weight of abdominal fat. The content of the crop was not affected by PD, whereas that of the proventriculus was lowest in the PD 2 group. Gizzard content was inversely related to PD. Pelleting reduced the length of the jejunum and ileum: which were shortened by about 15% with PDs 1 to 2, as compared to PD 0. The weight/length ratio of the jejunum and ileum tended to increase with increasing PD to a peak at PD 1·5, and to decrease thereafter.

6. Trypsin activity in the pancreas and amylase activity in the intestinal content were reduced by pelleting.

7. Chicks fed pelleted diets were less active: they ‘sat’ more and spent less time eating than their mash‐fed counterparts.     

Expression of matrix metalloproteinases in bovine luteal cells induced by prostaglandin F2α, interferon γ and tumor necrosis factor α

We recently demonstrated that luteal cells flow out from the ovary via lymphatic vessels during luteolysis. However, the regulatory mechanisms of the outflow of luteal cells are not known. Matrix metalloproteinases (MMPs) can degrade the extracellular matrix and basal membrane, and tissue inhibitors of matrix metalloproteinases (TIMPs) inhibit the activity of MMPs. To test the hypothesis that MMP expression in luteal cells is regulated by luteolytic factors, we investigated the effects of prostaglandin F2α (PGF), interferon γ (IFNG) and tumor necrosis factor α (TNF) on the mRNA expression of MMPs and TIMPs in cultured luteal cells. Luteal cells obtained from the CL at the mid-luteal stage (days 8–12 after ovulation) were cultured with PGF (0.01, 0.1, 1 μM), IFNG (0.05, 0.5, 5 nM) and TNF (0.05, 0.5, 0.5 nM) alone or in combination for 24 h. PGF and IFNG significantly increased the expression of MMP-1 mRNA. In addition, 1 μM PGF in combination with 5 nM IFNG stimulated MMP-1 and MMP-9 mRNA expression significantly more than either treatment alone. In contrast, IFNG significantly decreased the level of MMP-14 mRNA. The mRNA expression of TIMP-1, which preferentially inhibits MMP-1, was suppressed by 5 nM INFG. One μM PGF and 5 nM IFNG suppressed TIMP-2 mRNA expression. These results suggest a new role of MMPs: luteal MMPs stimulated by PGF and IFNG break down the extracellular matrix surrounding luteal cells, which accelerates detachment from the CL during luteolysis, providing an essential prerequisite for outflow of luteal cells from the CL to lymphatic vessels.     

Cloning,Expression of Clostridium perfringens α-toxin Gene and Preparation of its Antisera

One pair of primers had been designed and synthesized based on the α-toxin gene of Clostridium perfringens.The complete α-toxin gene fragment was amplified by polymerase chain reaction (PCR), and then was cloned into pGEM-T Easy vector to construct pGEM-T-α.Digested with EcoRⅠ and Hind Ⅲ, a fragment of 1125 bp was cloned into the expression plasmid vector pET-28a(+).The recombinant plasmid was transformed into the BL21(DE3)plys and induced by 1.0 mmol/L IPTG at 37 ℃.The expression product was found to be 46.1 ku as expected one identified by SDS-PAGE, and confirmed by Western blotting with Clostridium perfringens type A antisera, indicating similar reactivity with native α-toxin.Recombinant α-toxin protein was simultaneously found in culture supernatant, postsonic supertanant and inclusion bodies, most protein was expressed in inclusion bodies, which indicated recombinant α-toxin protein was expressed in the extracellular, periplasm and cytoplasm.Recombinant α-toxin protein in postsonic supertanant could not make mice die, indicating its non-toxicity.Toxin-antitoxin neutralization test showed that antisera of recombinant α-toxin protein were specific to α-toxin.Upon immunization of rabbit with the recombinant α-toxin protein, antisera with high antibody titer neutralizing 100 MLD toxin per 1 mL were prepared.     

Piloting the effectiveness of pig health education in combination with oxfendazole treatment on prevention and/or control of porcine cysticercosis,gastrointestinal parasites,African swine fever and ectoparasites in Angónia District,Mozambique

A community-based intervention combining health education (HE) and treatment of pigs for control of porcine cysticercosis (PC), gastrointestinal (GI) helminths, African swine fever (ASF) and external parasites was tested involving six villages of resource-poor smallholder pig farmers. Farmers and pigs of six rural villages were randomly allocated into group 1 (HE), which served as controls, and group 2 (HE?+?OFZ) pigs received a single oral dose of 30 mg/kg OFZ. Farmers were trained in pig health, housing and feeding. The proportion of farmers with confined pigs, the adoption rate of the introduced pig pen, the sero-prevalence of PC and ASF, the prevalence and intensity of GI nematodes and the prevalence of ectoparasites were measured at 9, 15 and 24 months after initiation and compared to a baseline survey to seek the effectiveness of the interventions. There was no clear effect of the intervention on the sero-prevalence of PC, but analysis of the rate of change in prevalence between the two groups showed significant effect with the rate of change to lower prevalence in the HE?+?OFZ group compared to the HE group. Although HE managed to improve the farmer’s knowledge in the control and prevention of ASF and ectoparasites, there was no significant reduction in the sero-prevalence of ASF and the prevalence of ectoparasites throughout the two-year period. The reported ineffectiveness of the intervention in this study suggested that more research is needed to develop more effective methods for controlling PC, ASF and pig parasites.     

Differential gene expression of antimicrobial peptides β defensins in the gastrointestinal tract of Salmonella serovar Pullorum infected broiler chickens

Salmonella enterica serovar Pullorum causes substantial mortality in chicks as well as results in persistent infection and vertical transmission in layer birds. An effective innate immune response in the early stages of infection could reduce bacterial colonization and mortality in chicks and persistency of infection in later stages. β Defensins (AvBDs) are now considered as one of the key components of innate immunity in avian species. In the present study, we quantified the mRNA expression levels of AvBDs (1–14) by real-time PCR in the gastrointestinal (GI) tissues (duodenum, jejunum, ileum and caecum) of 3-day-old broiler chicks after 24 h of oral infection with Salmonella Pullorum. Quantitative real-time PCR analysis revealed significant (P < 0.05) upregulation of AvBD3, 4, 5, 6 and 12 and a significant (P < 0.05) down regulation in the expressions of AvBD10, 11, 13 and 14 in one or few GI tissues, while no significant changes were observed for AvBD1, 2, 7, 8 and 9 gene expressions in any of the GI tissues investigated upon infection with S. Pullorum. Most substantial change in gene expression was found for AvBD5, being significantly (P < 0.01) upregulated in most of the GI tissues investigated. The differential expression levels of β defensins shed light on tailored innate immune response induced by S. Pullorum during the early stages of infection in chicks.     

Morphological features of the gastrointestinal tract of Garganey (Anas querquedula,Linnaeus 1758)—Oesophagus to coprodeum

The present work was aimed to study the gastrointestinal tract in Garganey from the oesophagus to coprodeum by the aid of the gross morphology, scanning electron microscopy and histology. At the level of the thoracic inlet, the oesophagus had a thick part which had numerous tortuous longitudinal folds carried transverse folds on them. Histologically, the thick part had thick epithelium and appeared more folded with more and larger glands. The mucosa of the proventriculus had mucosal folds appeared as a leaf-like with rounded tips. The openings of the proventriculus glands were projected with papillae on the epithelial surface. The gizzard lined with horizontal layer of cuticle which sent vertical parts of cuticle. The duodenal villi were pentagonal in shape with sharply pointed tips, while the jejunal villi were triangular with rounded ends, and the ileal villi were wide and pentagonal in shape with pointed tips. The mucosa of the duodenum, jejunum and ileum was thrown into villi lined by simple columnar epithelium with goblet cells with difference in the form, length, density and number. The apical points of the duodenal and ileal villi appeared pointed, while the jejunal villi had rounded ends. The jejunal villi were longer and numerous than the duodenal villi, but the ileal villi appeared with different length and less numerous. The mucosa of the caecum had numerous mucous glands which surrounded by circular crypts. The rectal mucosa had longitudinal folds and semilunar and circular crypts. The mucosa of the coprodeum had longitudinal folds surrounded by crypts.     

Expression of α1-acid glycoprotein and lipopolysaccharide binding protein in visceral and subcutaneous adipose tissue of dairy cattle

Adipose tissue is an endocrine compartment that plays an important role in immune defence by producing and releasing a wide range of proteins, including acute phase proteins (APPs). The liver is the main organ of APP synthesis, although extrahepatic production has also been reported. In the present study, expression of two APPs in dairy cattle, lipopolysaccharide binding protein (LBP) and α1-acid glycoprotein (AGP), was determined in four visceral (pericardial, mesenteric, omental and retroperitoneal) and three subcutaneous (withers, tail head and sternum) adipose tissue depots. mRNA expression was evaluated using qualitative and quantitative PCR, protein profiles were assessed by Western blot analysis and cellular localisation was determined by immunohistochemistry. The presence of LBP and AGP was demonstrated at mRNA and protein levels in all seven adipose tissue depots. Expression of AGP and LBP suggests that they may have roles as local and systemic inflammatory adipokines.     

Involvement of tannins and flavonoids in the in vitro effects of Newbouldia laevis and Zanthoxylum zanthoxyloïdes extracts on the exsheathment of third-stage infective larvae of gastrointestinal nematodes

The present study aimed at examining the possible role of tannins and flavonoids on the in vitro anthelmintic properties of the extracts of two plants from the southern area of Western Africa, i.e. Newbouldia laevis and Zanthoxylum zanthoxylo?des. Extracts of the two plants were prepared by use of acetone/water (70/30) and their anthelmintic activity was measured by use of the larval exsheathment inhibition assay (LEIA) applied on the abomasal species, Haemonchus contortus and the intestinal species Trichostrongylus colubriformis. Three concentrations of extracts were evaluated to examine the possible dose effect. In addition, the possible involvement of tannins and flavonoids was examined by comparing the levels of inhibition of larval exsheathment obtained with the same extracts, after of not addition of PVPP which forms complexes with these compounds. The results indicate significant effects with both plants and both nematode species. In the range of concentrations examined, the results were dose-dependent for N. laevis extracts but not for Z. zanthoxylo?des because the three doses applied provoked a similar highly significant inhibition whatever the tested dose. The use of PVPP indicated for both plant and nematode species, that tannins and flavonoids are involved partly in the effect but that some other biochemical compounds were also involved in both plants.     

Circadian and circannual rhythms of cortisol,ACTH, and α-melanocyte-stimulating hormone in healthy horses

Cosinor analysis was used to evaluate whether pituitary and adrenal hormones exhibit circadian rhythmicity in horses. The effect of season and animal age on their respective rhythms was also determined. In addition, the usefulness of evaluating cortisol rhythmicity for the diagnosis of pituitary pars intermedia dysfunction (PPID) was assessed. Serum cortisol concentrations (P < 0.01), but not plasma ACTH or α-melanocyte-stimulating hormone (α-MSH), showed a significant circadian periodicity in horses. An effect of season on hormone concentration was observed with plasma ACTH and α-MSH concentration greater in the fall and cortisol concentration greater in the spring (P < 0.001). Age did not affect cortisol rhythm, but it did blunt the variation in cortisol concentration in horses, similar to what has been previously reported to occur in aged people and dogs. In addition, our results suggest that clinically and diagnostically normal, non–PPID-affected horses commonly have a loss of cortisol diurnal rhythm. Therefore, measurement of circadian rhythm is not an appropriate diagnostic test for PPID.     

Effect of Freezing Rates and Supplementation of α-Tocopherol in the Freezing Extender in Equine Sperm Cryosurvival

This study was conducted to determine the impact of antioxidant (α-tocopherol) supplementation in the cryopreservation extender and three different freezing rates (FRs) on quality of post-thaw semen to elaborate a new protocol for stallion semen cryopreservation. Six ejaculates from each of four stallions were subjected to cryopreservation with a commercial extender (Gent, Minitub Iberia, Spain), without any supplementation (control) or supplemented with 2 mM α-tocopherol. The semen was exposed to three different freezing rates between 5°C and −15°C: slow (5°C/min), moderate (10°C/min), and fast (20°C/min). After thawing, the sperm viability (Sybr-14 and propidium iodide [PI]), mitochondrial membrane potential (MMP) (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine), lipid membrane peroxidation (LPO; C₁₁-BODIPY^581/591), and apoptosis status (fluorescein isothiocyanate–conjugated annexin V and PI) of the plasmatic membrane of each sample were determined by flow cytometry. For both extenders, the percentage of viable cells was higher for spermatozoa cooled at 5°C/min than at 10°C/min and 20°C/min (P ≤ .05). The FR of 20°C/min demonstrated a lower value of MMP than the FR of 5°C/min and 10°C/min (P ≤ .05). The α-tocopherol extender improved (P ≤ .05) post-thaw membrane LPO; however, it did not improved viability and the apoptosis status of the sperm plasmatic membrane after thawing. In conclusion, results clearly indicate that the cryosurvival of stallion spermatozoa is improved when FR of 5°C/min is used from 5°C to −15°C.     

Toluene Exposure Leads to a Change in Expression Patterns of β Defensins in the Mouse Tracheal Epithelium

Defensins are generally implicated in the quick resistance of epithelial surfaces to microbials; however, recent reports have indicated that defensins also have unknown purposes in relation to noninfectious diseases. In this study, the localization patterns of anti-microbial peptides, β defensins (BDs), in the tracheal epithelium of male C3H mice under exposure to toluene were analyzed by immunohistochemistry. Mice were exposed one to ten times to toluene for 30 min by nose-only inhalation. Expression of BDs was revealed by immunohistochemistry in serial sections of trachea after the final exposure. Expression of BD-1 was usually observed at almost the same levels in all exposure groups, and expression of BD-2 was observed in the control group; however, the signals for BD-2 decreased gradually with frequency of exposure. In the group exposed ten times, expression of BD-2 decreased to far lower than that of the control group. No expression of BD-3 was detected in any groups. Interestingly, expression of BD-4 increased to the maximum in the group exposed four times and decreased to a level lower than that of the control in the group exposed ten times. The results of the present study indicated that toluene gas might change the expression pattern of BDs in the tracheal epithelial cells. The oscillation of expression of BD-4 was quite characteristic and might contribute to morphological damage in on the epithelial cells.     

Isolation, characterization and establishment of an equine retinal glial cell line: a prerequisite to investigate the physiological function of Müller cells in the retina

Retinal Müller glial cells are of vital importance for maintaining a physiological environment within the retina. To this end, they provide highly specialized physiological properties to support neurons in structure, nutrition and metabolism. The purpose of this study was to isolate Müller cells from the equine retina, determine their characteristics and subsequently establish a stable equine Müller cell line (eqMC) that will provide a prerequisite for investigations on their physiological properties. Dissociated retinal cells were obtained from equine retinas by a papain digestion technique followed by trituration and a cell attachment method by which pure Müller cell cultures were achieved. Morphological examination was performed using phase-contrast microscopy, and further characterization of different subcultures was accomplished by immunocytochemistry. Cells of passage 1 showed distinct signals for glutamine synthetase and vimentin, whereas glial fibrillary acidic protein expression was almost absent. Characteristic expression patterns remained unaltered in all subcultures. Furthermore, cultured Müller cells stably expressed the microfilament alpha-smooth muscle actin, the proliferation marker Ki67 and the membrane channels Kir4.1 and aquaporin 4. The present study introduces the eqMC-7 that will facilitate studies investigating the physiological role of Müller cells within the equine retina.     

Effects of α-lipoic acid, vitamins E and C upon the heat stress in Japanese quails

Antioxidant vitamins are commonly used as colorant, preventive, sweetener, nutritive and antioxidant materials in food industry. Thus, many studies have been conducted in recent years. We have performed the present study to understand the effects of α-lipoic acid (ALA), vitamin C and E, added into rations of Japanese quails, on chronic heat stress. This study was carried out in five groups as: control, stress, ALA, vitamin E and vitamin C groups. Heat stress is applied to all groups except the control group. Superficial pectoral muscles tissues were used for biochemical determinations. During the research, it is found that heat stress exerted undesired effects such as increasing lipid peroxidation (LPO) level indicating oxidative stress in Japanese quails. In addition, superoxide dismutase (SOD) and catalase (CAT) enzyme activities and glutathione (GSH) amounts were studied to understand the behaviour of defence mechanism. While stress increased LPO ratio, it was determined that all antioxidant added into the ration decreases LPO significantly (p < 0.05). In addition, it was determined that all of antioxidants added into ration effectively restored SOD activity and LPO ratio which heat stress affected negatively. Interestingly, vitamin C did not adjust GSH ratio in contrast to vitamin E and ALA, where vitamin E and C did not perform any positive effect on heat stress decreased CAT activity. Finally, it can be thought that antioxidant vitamins relatively ameliorated these undesired affects caused by stress factors given.