Investigation on association and expression of ESR2 as a candidate gene for boar sperm quality and fertility

ESR2 is involved in oestrogen-related apoptosis in cell cycle spermatogenesis but their effects have not yet confirmed in pig. Therefore, this study was aimed to investigate the association of ESR2 polymorphism with sperm quality and boar fertility traits and to analyse the ESR2 mRNA and protein expressions in boar reproductive tissues. DNA samples from 203 Pietrain (PI) and 100 Pietrain × Hampshire (PIHA) pigs with records of sperm quality [sperm concentration (SCON), motility (MOT), semen volume (VOL), plasma droplet rate (PDR) and abnormal spermatozoa rate (ASR)] and fertility [non-return rate (NRR) and number of piglet born alive (NBA)] traits were available. A SNP in coding region of ESR2 g.35547A>G in exon 5 was associated with MOT and PDR in the PI and with SCON, VOL, MOT and PDR in PIHA population. For mRNA and protein expression study, a total of six boars were divided into two groups with group I (G-I) and group II (G-II) where G-I characterized for relatively a better sperm quality according to the mean of two groups. mRNA expression was higher in brain and testis than that in all parts of epididymis. Both qRT-PCR and western blot analysis revealed that the ESR2 gene expression and protein expression were significantly higher in testis collected from G-II compared with that of G-I boars. Moreover, ESR2 protein localization in germ cell, Leydig and Sertoli cells, epithelial cells and spermatozoa was remarkable, which indicated the important role of ESR2 in spermatogenesis process. These results might shed new light on the roles of ESR2 in spermatogenesis as candidate for boar fertility, but still the lack of association across populations should be considered.     

Dairy cow preference for a soft track surface

Abstract

Extract

As dairy herds get larger cows have to walk further, and foot care becomes more important. Lameness from bruising of the soles of the feet is a particular hazard (Tranter and Morris 1991 Tranter, W P and Morris, RS. 1991. A case study of lameness in three dairy herds. New Zealand Veterinary Journal, 39: 88–96. [Taylor &; Francis Online], [Web of Science ®] , [Google Scholar], especially on hard tracks with loose angular stones or grit (Clackson and Ward 1991 Clackson, DA and Ward, WR. 1991. Farm tracks, stockman's herding and lameness in dairy cattle. Veterinary Record, 129: 511–512.  [Google Scholar]. One way of reducing this problem might be to provide an overlay of woodchips on top of the normal hard track. This would provide a softer grit-free surface, which could be more comfortable for the cows.     

The distribution of taste buds in Garra rufa

The distribution and relative frequency of external taste buds (TB) of Garra rufa were studied. TB of Garra rufa were observed on different body location (lips, lateral and ventral areas, forehead, operculum and dorsal-pelvic-pectoral-anal fins). TB are at the highest frequency in lips, forehead, pectoral and anal fins. These structures are moderate as a number in operculum, La2 (between pelvic and anal fins) areas, and then decreased a few in other areas.     

Distribution and ontogeny of gastrin- and serotonin-immunoreactive cells in the proventriculus of developing chick, Gallus gallus domestica

The ontogeny and distribution of gastrin- and serotonin-immunoreactive (IR) cell in the proventriculus of chicks (Gallus gallus domestica, n = 60) in different growth periods was examined immunohistochemically using antisera specific to gastrin and serotonin. Gastrin and serotonin-IR cells were detected in chick proventriculus. Gastrin-IR cells were first evident after 12 days of incubation in lamina epithelialis and compound glands, while serotonin-IR cells were observed only in compound glands at that same time. Gastrin-IR and serotonin-IR cells increased in frequency on incubation day 14 and 16, respectively. Towards the end of incubation, gastrin- and serotonin-IR cell numbers decreased. In adult chicken, both IR cells were present but not lower numbers. The observations demonstrate the presence of gastrin- and serotonin-IR cells in the proventriculus of developing chicks in temporally changing frequencies.     

Histological and histochemical characterization of the mucosa of the digestive tract in flower fish (Pseudophoxinus antalyae)

The wall of the digestive tract is composed of the tunica mucosa, tunica muscularis and tunica serosa. Lamina-submucosa separation or any glands were not observed in tunica mucosa. Goblet cells were determined to constitute a much larger reserve at digestive tract mucosa. Histochemical analysis of the intestine of flower fish (Pseudophoxinus antalyae) showed that gastrointestinal mucous content included sulphate-esters and/or carboxylic [Alcian blue (AB) 0.06+], glycogene and/or oxidable dioles [periodic acid/Schiff+ (PAS+)], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS) and strong acid sulphated [Aldehyde fuchsin+ (AF+)] glycoproteins (GPs). Except these mucosubstances to lower densities, densely sulphate (AB pH 2.5+), O-sulphate esters (AB pH 1+) strong and weak sulphated (AB 0.3 M+), GPs were also determined.     

Investigation into association and expression of PLCz and COX-2 as candidate genes for boar sperm quality and fertility

Phospholipase C zeta (PLCz) and cyclooxygenase isoenzyme type 2 (COX-2) are important in spermatogenesis, but their effect has not yet confirmed in pigs. Therefore, this study was aimed to analyse their association with sperm quality and fertility and to identify the mRNA and protein expression in boars reproductive tissues. DNA samples from 231 Pietrain (PI) and 109 Pietrain × Hampshire (PIHA) pigs with records of sperm quality [sperm concentration (SCON), motility, semen volume, plasma droplet and abnormal spermatozoa rate] and fertility (non-return rate and number of piglet born alive) traits were available. A SNP in non-coding region of PLCz g.158 A > C was associated with SCON (p < 0.05) in PIHA population while the polymorphism of COX-2 g.68 G > A in 3' UTR was not associated with any traits. For mRNA and protein expression study, a total of six boars were divided into two groups with G-I and G-II, where G-I was characterized for relatively better sperm quality. Both genes expressed higher in reproductive tissues compared with non-reproductive tissues. Phospholipase C zeta mRNA expressed higher in testis (p < 0.01), all parts of epididymis and spermatozoa from G-I, while COX-2 expressed higher in testis (p < 0.05), head and body of epididymis (p < 0.01), and spermatozoa from G-II boar. Both proteins were localized in Leydig cells and spermatozoa. These results might shed light on roles of these genes in spermatogenesis as candidate for boar sperm quality and fertility, but still the lack of association across populations should be considered.     

The distribution of external taste buds in flower fish (Pseudophoxinus antalyae)

Purpose of this study was to determine the distribution of external taste buds of flower fish (Pseudophoxinus antalyae) on different body locations (lip, lateral, ventral and dorsal areas, dorsal-pelvic-pectoral-anal fins). In this species, it was found that the density of taste buds was greater on lip epithelium than that of lateral (between pectoral-anal, anal-caudal fins), ventral (between pelvic and anal fins) and dorsal (close to the head) locations. It was observed that the shape of taste bud changed depending upon the epithelium thickness and it was found that some of these structures protruded towards to the exterior of epithelium.     

The histology and histochemical aspects of gills of the flower fish, <Emphasis Type="Italic">Pseudophoxinus antalyae</Emphasis>

Branchial epithelium of Pseudophoxinus antalyae was lined by both a thick stratified epithelium lining gill arches, gill rakers and primary filaments and a thin epithelium lining the lamellae. Mucous, chloride and rodlet cells, interspersed between pavement cells, were present in the branchial epithelium. With histochemical procedures for the characterization of glycoconjugates, mucous cells showed a strong positive reaction with Periodic acid-Shiff and Alcian Blue at pH 2.5, although with Alcian Blue at pH 0.5 and pH 1.0 the reaction was much weaker. When the combined Alcian Blue (pH 2.5) – Periodic acid-Shiff reaction was performed, most mucous cells were stained purple, whereas by the combined Aldehyde Fuchsin/Alcian Blue (pH 2.5), most cells showed a positive reaction only to Aldehyde Fuchsin. Methylation/ Alcian Blue (pH 2.5) and Methylation/ Saponification/ Alcian Blue (pH 2.5) methods showed the presence of sulphated and carboxylated glycoconjugates in mucous cells. Mucous cells were also detected to stain all metachromatically with Toluidine Blue.     

Blood pressure and electrocardiographic effects of acepromazine in anaesthetized horses

Acepromazine, a phenothiazine tranquilizer, causes hypotension in standing horses ( Parry et al. 1982 ). However, a retrospective study ( Taylor & Young 1993 ) showed that acepromazine pre‐anesthetic medication did not affect arterial blood pressure (MAP) in anaesthetized horses. This study examined the effects of acepromazine on MAP during romifidine–ketamine–halothane anaesthesia in horses anaesthetized for various surgical procedures. Forty‐four horses were allocated by block randomization to groups A and B. Group A received acepromazine 0.05 mg kg^?1 IM 30 minutes before induction of anaesthesia, group B did not. All horses received romifidine 0.1 mg kg^?1 IV 5 minutes before anaesthesia was induced with diazepam 0.05 mg kg^?1 and 2.2 mg kg^?1 ketamine IV. The horses' trachea were intubated and horses breathed 50% oxygen and 50% nitrous oxide plus halothane (concentration adjusted as required clinically) from a circle breathing system. Nitrous oxide was discontinued after 10 minutes and analgesics, flunixin 1.1 mg kg^?1 and either morphine 0.1 mg kg^?1 or butorphanol 0.05 mg kg^?1 (matched for horses undergoing the same procedure) administered IV. The facial or dorsal metatarsal artery was catheterized for direct measurement of MAP (every 10 min) and withdrawal of blood for gas analysis (every 30 min). The electrocardiogram (ECG) was monitored continuously with a 10 seconds printout obtained every 10 minutes. Intermittent positive pressure ventilation (IPPV) was instigated if PaCO₂ exceeded 9.3 kPa (70 mm Hg). Dobutamine was infused (1.0–5.0 kg^?1minute^?1) if MAP < 58 mm Hg and was continued until MAP > 70 mm Hg. Mean age, weight and duration of anaesthesia were compared between the groups using a t‐test for independent samples. Gender distribution and numbers of horses requiring IPPV or dobutamine were compared between groups using a chi‐squared test (with Yates correction). To compare MAP over time, the area under the curve (MAPAUC) was calculated and compared between groups using a t‐test. Horses receiving dobutamine were excluded from MAPAUC and MAP comparisons. The ECG printouts were examined for arrhythmias. There were no significant differences between groups (p > 0.05). Group A contained three stallions, 10 geldings and nine mares, aged 6.3 years (range 0.75–18). Group B comprised eight stallions, 11 geldings and three mares aged 7.3(1–16) years. Duration of anaesthesia was group A 97 (50–140) minutes, group B 99 (50–160) minutes. Eight horses in group A and three in group B required IPPV. Nine horses in group A and four in group B received dobutamine. Mean arterial pressure ranged from 60 to 128 mm Hg in group A and 58–96 mm Hg in group B. Mean MAPAUC was 5941 mm Hg minute^?1 in group A, in B 6000 mm Hg minute^?1. Atrial pre‐mature complexes were recorded from one horse in group B. No other arrhythmias were detected. Although MAP was lower in the acepromazine group, this appeared unlikely to cause a clinical problem. The incidence of arrhythmias was too low to determine the influence of acepromazine in this study.     

The effects of levamisole on oxidative stress induced by copper intoxication in broilers

Abstract

AIM: To determine the effects of Cu and levamisole on concentrations of Cu and Fe in plasma and liver, and the effects of levamisole on lipid peroxidation induced by Cu intoxication in broiler chickens.

METHODS: In a 2×4 factorial study, 80 one-day-old Ross PM3 broiler chicks were fed diets for 21 days containing either 8 mg/kg Cu (Low Cu) or 250 mg/kg Cu (High Cu) and were treated with 0 (L0), 4 (L4), 8 (L8) or 16 (L16) mg/kg bodyweight levamisole per day from Day 7 of the study, on three consecutive days in their drinking water. This treatment was repeated three times, at 3-day intervals. On Day 21, blood samples were collected from each bird for analysis of concentrations of Cu, Fe and malondialdehyde, and activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), superoxide dismutase, catalase and glutathione peroxidase (GSH-Px). The birds were killed and liver samples collected for analysis of Cu and Fe.

RESULTS: Mean concentrations of Cu and Fe in plasma, and Cu in liver, were increased overall in the High Cu groups compared with the Low Cu groups (p<0.001). Compared with the L0 treatment group on the High Cu diet, treatments L4, L8 and L16 decreased concentrations of Cu in plasma, and L8 and L16 increased concentration of Cu in liver (p<0.05). Mean activities of AST and ALT were increased in untreated birds (L0) fed the High compared with Low Cu diets (p<0.01). In birds receiving the High Cu diet, treatments L4 and L8 decreased activities of AST, and L4 and L16 decreased activity of ALT, compared with L0 (p<0.05). The High Cu diet induced an oxidative stress characterised by increased mean concentrations of malondialdehyde and decreased activities of superoxide dismutase, catalase and GSH-Px (p<0.001). Concentration of malondialdehyde, and activities of superoxide dismutase and catalase were not changed following levamisole treatment in birds on the High Cu diet, and activity of GSH-Px was decreased by the L4 and L8 treatments compared with the L0 group.

CONCLUSIONS AND CLINICAL RELEVANCE: The results of the study suggest that treatment with levamisole might alleviate the harmful effects of Cu on the liver, as demonstrated by decreased activities of AST and ALT induced by a diet containing 250 mg/kg Cu.