Immunohistochemical Detection of Mycoplasma agalactiae in Formalin‐Fixed,Paraffin‐Embedded Tissues from Naturally and Experimentally Infected Goats

Samples from the mammary tissue of 14 lactating goats (12 naturally infected and two experimentally infected) were examined for the presence of Mycoplasma agalactiae. A monoclonal antibody (5G12) was applied to formalin‐fixed, paraffin‐wax‐embedded sections and labelled by the avidin–biotin peroxidase complex (ABC) method. Histological examination of tissue sections revealed strong immunoreactivity in all animals included in the study. Mycoplasma agalactiae antigen was mainly detected in the cellular debris at the periphery of purulent exudates present within lactiferous sinuses, and lactiferous and interlobular ducts. In addition, M. agalactiae organisms appeared in the cytoplasm of the epithelium of ducts, and in infiltrating macrophages and neutrophils within the ducts, alveoli, interstitial tissue and regional lymph node sinuses. It is concluded that this monoclonal antibody‐based immunohistochemical technique is an efficient and specific method for the post‐mortem detection of M. agalactiae in cases of clinical mastitis as well as being a useful tool for the study of the route of infection and cellular types involved during mastitis caused by this organism.     

CHEMICAL AND BIOLOGICAL PROPERTIES OF TWO NEW HERBICIDES—IOXYNIL AND BROMOXYNIL

Summary. The preparation of ioxynil, bromoxynil, and their salts is described, and information on solubilities and stability to storage is summarized. Although the toxicology of the herbicides is to be reported in greater detail, a preliminary statement is made here. Evidence of herbicidat activity under glasshouse conditions is indicated briefly, and supports the conclusion that both compounds are effective when applied to the foliage of a wide range of dicotyledon weed species. Seedlings of some weed species resistant to the phenoxy alkanoic acids are controlled under glasshouse conditions at doses as low as 0.125 lb/ac, and ioxynil has a wider range of activity than bromoxynil at these low doses. Graminaceous species tolerate 4–8 lb/ac of both herbicides without injury, and certain leguminous crops tolerate one or other herbicide at doses of 0.5–0.75 lb/ac. The contact action of the herbicides is rapid, there are also slower systemic effects, and seed germination is inhibited. In susceptible species the level of post-emergence activity is shown to be influenced by the growth stage of the weed, the distribution of herbicide on the foliage, and environmental factors of which light intensity appears to be most important.
Propriétés chimiques et biologiques de deux nouveaux herbicides: ioxynil et bromoxynil     

Studies on the potential use of Pasteuria penetrans as a biocontrol agent of root-knot nematodes (Meloidogyne spp.)

Some aspects of the interaction of the bacterial parasite Pasteuria penetrans and the root-knot nematode ( Meloidogyne spp.) were investigated in laboratory and pot experiments. The variable spore attachment on juveniles exposed to water suspensions of the bacterium is probably attributed to differential susceptibility of biotypes within a heterogeneous Meloidogyne population. The relationship between spore concentration and attachment level is not linear over a range of spore dosages, indicating that even at very high spore concentrations the number of spores capable of attachment may not be present in excess and it is difficult to ensure sufficient numbers of spores to ensure infection will attach to all nematodes. Attempts to apply the bacterium in conditions such as might occur in seedbeds did not suppress nematode multiplication after transplanting in nematode-infested soil, indicating that the only effective application method is a thorough spore distribution in the planting sites. Two major constraints were revealed: high levels of spore attachment to juveniles does not always guarantee a significant reduction of egg laying and this is greatly influenced by the Meloidogyne biotype. Furthermore, the cumulative effect of the parasite in reducing Meloidogyne populations over several crop cycles was less than expected as the bacterium reduced intra-specific competition for the food supply and the less damaged root enabled many nematodes to survive.     

Feeding fodder beet (Beta vulgaris L.) with either barley straw or pasture silage to non-lactating dairy cows

AIMS: To determine the suitability of diets containing either approximately 85% fodder beet (Beta vulgaris L.) with barley straw or 65% fodder beet with pasture silage when fed to non-lactating dairy cows, by measuring intakes, digestibility, rumen function including microbial growth, and N excretion.

METHODS: Holstein-Friesian cows fitted with permanent rumen fistulae were fed either 65% fodder beet with pasture silage (Silage; n=8) or 85% fodder beet with barley (Hordeum vulgare L.) straw (Straw; n=8) in an indoor facility over a 9-day period, for measurement of intakes, digestibility, rumen function and urine production. The cows were adapted to the diets over 2 weeks before the indoor measurements. Feed was available for about 6 hours/day, as practiced commercially for wintering non-lactating cows.

RESULTS: Five cows fed the Straw diet had to be removed from the trial because of acute acidosis; four on Day 1 of the measurement period and one on Day 7. One cow allocated to the Silage diet refused to eat fodder beet bulbs and was also removed from the trial. Two cows fed the Silage diet were also treated for acidosis. DM intakes were lower with the Straw than Silage diets (6.4 (SE 0.4) vs. 8.3 (SE 0.5) kg/day) and organic matter (OM) digestibility was lower with the Straw than Silage diets (77 (SE 1) vs. 83 (SE 1) g/100g). The N content of the two diets was 1.14 and 1.75?g/100?g DM and there was a net loss of N by cows fed the Straw diet (?22.7 (SE 7) g/day). Rumen microbial N production was much lower in cows fed the Straw than the Silage diet (6.6 (SE 1.3) vs. 15.8 (SE 0.7) g microbial N/kg digestible OM intake). Concentrations of ammonia in rumen liquid collected on Days 5–6 were below detection limits (<0.1?mmol/L) in 36/48 (75%) samples collected from cows fed the Straw diet and in 27/48 (56%) cows fed the Silage diet. Mean urinary N excretion was lower in cows fed the Straw than the Silage diet (52.0 (SE 5.8) vs. 87.7 (SE 5.9) g/day).

CONCLUSION AND CLINICAL RELEVENCE: An over-wintering diet for dry cows comprising about 65% fodder beet with 35% pasture silage provided adequate nutrition, although there was some risk of acidosis. In contrast, the diet containing about 85% fodder beet with barley straw resulted in lower DM intakes, poor rumen function, negative N balance so that both nutrition and welfare were compromised.     

Spermatic and oxidative profile of domestic cat (Felis catus) epididymal sperm subjected to different cooling times (24, 48 and 72 hours)

Cooling stored epididymal samples for several days allows facilities to transport and process genetic material post‐mortem. Improvements to this practice allow the preservation of sperm from domestic cats, which are the ideal study model for wild felids. However, the modifications in spermatic features and the oxidative profile are not fully understood in cats. This information is necessary for the development of biotechniques, such as new extenders for cryopreservation. Therefore, the purpose of this study was to evaluate the spermatic and oxidative profile in samples from the epididymal cauda of domestic cats cooled at 5°C for 24, 48 and 72 hr. Spermatozoa were collected from the epididymis cauda. Evaluations consisted of computer‐assisted sperm analysis (CASA), plasma membrane integrity (eosin/nigrosin), acrosome integrity (fast green/rose bengal), sperm morphology, sperm DNA integrity (toluidine blue), mitochondrial activity (3′3 diaminobenzidine), activity of the antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD), measurement of lipid peroxidation (TBARS) and protein oxidation. A decrease in sperm motility parameters was observed after 72 hr of cooling (i.e. total and progressive) with a higher percentage of minor (37.7 ± 6.3%) and total defects (53.4 ± 6.3%). Additionally, a decrease in high mitochondrial activity (Class I: 16.6 ± 2.2%) occurred after 72 hr. The decrease in motility rates after a long cooling time probably was caused by the increase in sperm abnormalities. A long cooling time causes cold shock and mitochondrial exhaustion, but there was no observed change with the oxidative stress condition. Therefore, cat epididymal sperm stored at 5°C appear to maintain a high quality for up to 48 hr of cooling time.     

A fast,low‐cost and efficient method for the diagnosis of sperm DNA fragmentation in several species

Sperm DNA fragmentation is a condition that interferes directly in the reproductive efficiency. Currently, there are several methods for assessing the sperm DNA integrity, such as Alkaline Comet, TUNEL and Sperm Chromatin Structure Assay. However, many of these techniques are laborious and require high‐precision equipment. Thus, the development of new techniques can optimize the evaluation of sperm DNA damage. Therefore, the aim of this study was to standardize the toluidine blue (TB) stain technique for the analysis of DNA fragmentation of dog, cat, bull, stallion and ram spermatozoa. For this purpose, we used six animals of each specie (n = 30), in reproductive age. Sperm was collected by different methods according to the particularities of each species, and such samples were divided into two aliquots: a sperm sample was kept at 5°C (considered as intact sperm DNA), and the remaining samples were submitted to the induction of DNA fragmentation by exposure to ultraviolet light for 4 hr. Samples were then mixed with the intact sample to obtain known and progressive proportions of sperm with fragmented DNA (0%, 25%, 50%, 75% and 100%). Semen smears were performed and subjected to staining with TB. Blue‐stained spermatozoa were considered to have DNA fragmentation. We observed high linear regression coefficients between the expected proportion of damaged DNA and the results of TB for dog, cat, ram, bull and stallion samples. In conclusion, TB stain was considered a fast and effective technique for the study of spermatozoa DNA in several species.     

多年生人工草地矮生嵩草(Kobresia humilis)种群空间格局与种内关联性

植物种群空间分布格局不仅反映植物的空间分布特点,还有助于研究者了解植物利用资源的现状和生存能力。本研究以矮生嵩草(Kobresia humilis)为研究对象,按株丛斑块面积将矮生嵩草分为小斑块级株丛(0～30 cm²)、中斑块级株丛(30～80 cm²)和大斑块级株丛(>80 cm²)3个株丛级,采用点格局分析的g函数统计方法,分析了矮生嵩草各级株丛的空间格局及关联性。结果表明：矮生嵩草种群中小斑块级株丛占比高于中斑块、大斑块级株丛占比,种群中幼苗充足,正处于稳定的增长阶段;矮生嵩草小斑块、中斑块、大斑块级株丛在小尺度范围内表现为聚集分布,随着尺度的增大,所有株丛斑块的聚集程度减弱,逐渐趋向于随机分布;小斑块与中斑块、大斑块的矮生嵩草株丛在小尺度内表现为正关联。斑块聚集分布和相互共存是矮生嵩草的一种生存策略。     

Occurrence of viral haemorrhagic septicaemia in rainbow trout Salmo gairdneri Richardson reared in sea-water

Abstract. Viral haemorrhagic septicaemia (VHS) is reported for the first time in sea-water cultured rainbow trout. Heavy mortalities with typical signs and lesions A VHS virus (serotype 1) was isolated from the diseased fish. The mortalities were caused only by the VHS virus and 80 days post transfer of trout to sea-water the mortalities reached 85%, of the initial population.
The disease was experimentally transmitted to rainbow trout, both in sea-water 3·10⁴ pfu/ml of virus or by intramuscular injection of various doses of VHS₁ (7·10¹ 7·10⁴ or 7·10⁴ pfu per fish). Death occurred in all infected groups and started earlier in sea-water. Typical signs of VHS were observed in moribund fish. Viral multiplication was demonstrated to have occurred in fish organs.