Apoptosis in the Antral Follicles of Swamp Buffalo and Cattle Ovary: TUNEL and Caspase-3 Histochemistry

The present study was carried out to investigate the pattern of apoptosis in the healthy antral and atretic follicles of Philippine swamp buffaloes (BU) in comparison with Holstein-Friesian (HF) cows. Paraffin sections of healthy follicles and various stages of atretic follicles were stained using the terminal deoxynucleotidyl transferase (TdT)-mediated biotinylated deoxyuridine triphosphates (dUTP) nick end-labelling (TUNEL) method to detect DNA fragmentation and cleaved caspase-3 antibody to detect cells committed to undergo apoptosis. Five equidistant areas of a follicle were counted for the presence of TUNEL- and caspase-3-positive cells. Healthy follicles of BU and HF contained no TUNEL-positive cells in the granulosa and theca layer but showed some caspase-3 positivity. The granulosa layer of advanced atretic follicles showed a significantly higher frequency of caspase-3 positivity than the healthy and early atretic follicles in both breeds. The frequency of caspase-3-positive cells of BU was significantly higher than HF in the granulosa layer of healthy, early atretic and advanced atretic follicles. In the theca interna layer, BU and HF showed a significantly lower and higher frequency of TUNEL-positive cells in the late atretic follicles compared with advanced atretic follicle, respectively. However, the frequency of caspase-3-positive cells of both BU and HF in the late atretic follicles was significantly higher than the advanced atretic follicles in the theca interna layer. These results indicate that caspase-3 and DNA fragmentation is involved in the buffalo ovarian apoptotic process.     

Cell Proliferation in the Atretic Follicles of Buffalo and Cattle Ovary

The present study was carried out to describe the proliferative activity of granulosa and theca cells in healthy antral and atretic follicles of Philippine buffaloes (BU) and Holstein-Friesian (HF) cows. Paraffin sections of ovary were immunostained with mouse monoclonal antibody to proliferating cell nuclear antigen (PCNA). Then the follicles were classified into healthy and various stages of atretic follicles. The granulosa layer of healthy follicles had a significantly higher frequency of PCNA-positive cells than the early and advanced atretic follicles in both breeds. In the theca interna, significantly reduced populations of the PCNA-positive cells were found in both breeds as atresia progressed. Moreover, HF had significantly higher PCNA-positive cells in the theca interna of healthy, early atretic and advanced atretic follicles than BU. A reduction of PCNA-positive cells during atresia was also noted in the theca externa in both animals although differences were not significant. The results of the present work suggest that the proliferative activity of granulosa and theca cells decreases in association with follicular atresia in the BU similar to HF. Furthermore, a significantly deficient cell proliferative activity of theca interna was found in BU compared with HF.     

Ergovaline does not alter the severity of ryegrass staggers induced by lolitrem B

AIM: To investigate a possible interaction between lolitrem B and ergovaline by comparing the incidence and severity of ryegrass staggers in sheep grazing ryegrass (Lolium perenne) containing lolitrem B or ryegrass containing both lolitrem B and ergovaline.

METHODS: Ninety lambs, aged approximately 6 months, were grazed on plots of perennial ryegrass infected with either AR98 endophyte (containing lolitrem B), standard endophyte (containing lolitrem B and ergovaline) or no endophyte, for up to 42 days from 2 February 2010. Ten lambs were grazed on three replicate plots per cultivar. Herbage samples were collected for alkaloid analysis and lambs were scored for ryegrass staggers (scores from 0–5) weekly during the study. Any animal which was scored ≥4 was removed from the study.

RESULTS: Concentrations of lolitrem B did not differ between AR98 and standard endophyte-infected pastures during the study period (p=0.26), and ergovaline was present only in standard endophyte pastures. Ryegrass staggers was observed in sheep grazing both the AR98 and standard endophyte plots, with median scores increasing in the third week of the study. Prior to the end of the 42-day grazing period, 22 and 17 animals were removed from the standard endophyte and AR98 plots, respectively, because their staggers scores were ≥4. The cumulative probability of lambs having scores ≥4 did not differ between animals grazing the two pasture types (p=0.41).

CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence for ergovaline increasing the severity of ryegrass staggers induced by lolitrem B. In situations where the severity of ryegrass staggers appears to be greater than that predicted on the basis of concentrations of lolitrem B, the presence of other tremorgenic alkaloids should be investigated.     

基于自适应逆控制技术的连续流粮食干燥系统研究

连续流粮食干燥过程是一个非线性大滞后多变量的过程,要精确控制粮食出口的终水分比较困难。本文设计一种新的连续流粮食干燥过程的控制方法,对连续流粮食干燥过程中的多个影响因素进行研究,分析粮食干燥自控系统在设计时所必须选择的可测干扰、数字变量、控制变量和被控变量等,并且提出基于自适应逆控制技术的连续流粮食干燥系统的离散化模型,主要针对工业控制对象中存在的时变和时滞问题,推导出该连续流粮食干燥控制系统的预测模型。通过实验对实际系统进行验证,结果证明本系统具有较好的鲁棒性和稳定性。     

Polyglandular endocrinopathy type II (Schmidt's syndrome) in a Dobermann pinscher

A three‐year‐old, female neutered, Dobermann pinscher was presented for investigation of lethargy, episodic collapse, ataxia and myxoedema. Primary hypothyroidism and primary cortisol‐deficient hypoadrenocorticism were diagnosed based on history, physical examination and compatible hormonal analysis. Increased serum concentrations of thyroglobulin autoantibodies and 21‐hydroxylase autoantibodies indicated an immune‐mediated aetiology. The case was complicated by lymphadenopathy with hand‐mirror lymphocytes, classically identified in lymphoma. A polymerase chain reaction test for antigen receptor rearrangement indicated polyclonality and therefore reactive lymphadenopathy. The dog's clinical signs resolved following introduction of levothyroxine and prednisolone. Prioritising the problem‐based approach in this case facilitated the diagnosis of hypoadrenocorticism in addition to hypothyroidism due to the persistence of clinical signs despite thyroxine replacement. Importantly, atypical adrenal gland dysfunction was not misinterpreted as inadequate therapeutic response to thyroxine supplementation. The observation that polyglandular endocrinopathy type II can occur in dogs suggests that in dogs with a suboptimal response to treatment for hypothyroidism or hypoadrenocorticism comorbid endocrinopathies should be investigated.     

Carbon isotopic ratios of wetland and terrace soil sequences in the Maya Lowlands of Belize and Guatemala

This article provides new data and synthesizes earlier findings on the carbon isotope ratios of the humin part of soil organic matter from a range of sites in the central Maya Lowlands. Changes down the soil profile in carbon isotope ratios can provide an important line of evidence for vegetation change and erosion over time, especially in well dated aggrading profiles. Research thus far has provided substantial evidence for significant inputs from C₄ vegetation in buried layers from the Ancient Maya periods in depositional soils but equivocal evidence from sloping soils. We present new findings from soil profiles through ancient Maya wetland fields, upland karst wetlands, ancient Maya aguadas (reservoirs), and ancient Maya terraces. Most of the profiles exhibited δ¹³C enrichment greater than the 2.5–3‰ typical from bacterial fractionation. Seven of nine ancient Maya wetland profiles showed δ¹³C enrichment ranging from 4.25 to 8.56‰ in ancient Maya-dated sediments that also contained phytolith and pollen evidence of grass (C₄ species) dominance. Upland karst sinks and ancient reservoirs produced more modest results for δ¹³C enrichment. These seasonal wetland profiles exhibited δ¹³C enrichment ranging from 1 to 7.3‰ from the surface to ancient Maya-period sediments. Agricultural terraces produced mixed results, with two terraces having substantial δ¹³C enrichment of 5.34 and 5.66‰ and two producing only equivocal results of 1.88 and 3.03‰ from modern topsoils to Maya Classic-period buried soils. Altogether, these findings indicate that C₄ plants made up c. 25% of the vegetation at our sites in the Maya Classic period and only a few percent today. These findings advance the small corpus of studies from ancient terraces, karst sinks, and ancient wetland fields by demonstrating substantial δ¹³C and thus C₄ plant enrichment in soil profile sections dated to ancient Maya times. These studies are also providing a new line of evidence about local and regional soil and ecological change in this region of widespread environmental change in the Late Holocene.     

Cryopreservation of Piau‐Breed Wild Boar Sperm: Assessment of Cooling Curves and Centrifugation Regimes

This study aimed to assess the effects of different cooling curves and centrifugation regimes used in cryopreservation protocols on the post‐thaw viability of Piau‐breed wild boar (Sus scrofa) sperm using in vitro assessment tests. Two centrifugations (800  g for 10 min and 2400  g for 3 min) and two cooling curves (conventional cooling using nitrogen vapour – freezing 1 and automated cooling using a programmed freezing machine – freezing 2) were tested. Therefore, the treatments were divided into M3 – centrifugation at 2400  g for 3 min and freezing 2; M10 – centrifugation at 800  g for 10 min and freezing 2; R3 – centrifugation at 2400  g for 3 min and freezing 1; and R10 – centrifugation at 800  g for 10 min and freezing 1. No significant differences (p > 0.05) between treatments occurred post‐thawing regarding the total sperm motility means recorded. The mean values of the different treatments were not different from each other regarding the supravital staining (SV), hypo‐osmotic test (HO), sperm–egg binding assay or sperm morphology. This study showed that both the cooling curve and the centrifugation regime affected the quality of post‐thaw sperm, and centrifugation for shorter times and cooling curves using automated cooling are the most suitable for minimizing sperm injury.     

Basic mammalian bone anatomy and healing.

The goal of any method of fracture repair should be the early return to function of the patient with minimum postoperative morbidity. This is accomplished most optimally by having a basic understanding of the biology of bone healing and by being familiar with the musculoskeletal system of the species before attempting fracture repair. Applying the fundamental principles of mammalian bone anatomy and physiology to the bird ensures the best prognosis possible and minimizes postoperative complications in the avian patient.