Clinical and pathological observations in English cocker spaniels with primary metabolic vitamin E deficiency and retinal pigment epithelial dystrophy

Fifteen English cocker spaniels with confirmed vitamin E deficiency were examined physically, ophthalmologically and neurologically. Eleven of them had clinical signs of neurological dysfunction which included ataxia, proprioceptive deficits, abnormal spinal reflexes and muscle weakness. In the two dogs examined histopathologically there was central neuronal fibre degeneration with prominent neuroaxonal dystrophy, particularly within the sensory relay nuclei of the brainstem, and one of the dogs had severe intestinal lipofuscinosis.     

Hypovirulence-Associated Double-Stranded RNA from Sclerotinia homoeocarpa Is Conspecific with Ophiostoma novo-ulmi Mitovirus 3a-Ld

ABSTRACT The nucleotide sequence of the hypovirulence-associated double-stranded RNA (dsRNA) in hypovirulent isolate Sh12B of Sclerotinia homoeocarpa, the causal agent of dollar spot of turf grass, was determined. This large dsRNA (L-dsRNA) is 2,632 bp long and is A and U rich (61.0% A+U residues). One strand of this dsRNA contains an open reading frame (ORF) with the potential to encode a protein of 720 amino acids. This ORF contains 12 UGA codons, predicted to encode tryptophan in ascomycete mitochondria, and has a codon bias typical of mitochondrial genes, which is consistent with a mitochondrial localization of this dsRNA. The amino acid sequence contains conserved motifs typical of RNA-dependent RNA polymerases (RdRps). Sequence analyses of the nucleotide and RdRp-like protein revealed that the L-dsRNA is homologous with previously characterized mitochondrial viruses and dsRNAs from other phytopathogenic fungi, and shares 92.4% nucleotide and 95.1% amino acid sequence identities with the Ophiostoma novo-ulmi mitovirus 3a-Ld from Ophiostoma novo-ulmi, the causal agent of Dutch elm disease. The results indicate that these two dsRNAs are conspecific. This is the first report that a hypovirulence-associated dsRNA virus naturally occurs in two taxonomically distinct fungi, and indicates that horizontal transmission of this dsRNA virus may have occurred between these fungi.     

Quantifying Aphanomyces euteiches in Alfalfa with a Fluorescent Polymerase Chain Reaction Assay

ABSTRACT A polymerase chain reaction (PCR) assay using a set of specific primers and a dual-labeled probe (TaqMan) was developed to quantify the amount of Aphanomyces euteiches DNA in alfalfa plants exhibiting varying levels of disease severity. The study included isolates of race 1 and race 2 of A. euteiches. The assay also discriminated between alfalfa populations for resistance based on analysis of DNA extracted from bulked plant samples. Analysis of individual plants and bulked plant samples of standard check populations with both pathogen isolates resulted in Spearman rank correlations between pathogen DNA content and disease severity index ratings that were greater than 0.75 and highly significant (P < 0.0005). In experiments with a race 1 isolate, the amount of pathogen DNA present in the resistant check WAPH-1 was significantly less than in the susceptible check Saranac. In experiments with a race 2 isolate, the amount of pathogen DNA in the resistant check WAPH-5 was significantly less than in either of the susceptible checks, Saranac and WAPH-1. Discrimination between commercial cultivars based on quantitative PCR analysis of bulked plant samples was similar to classification based on visual assessment of disease severity.     

Novel method for the production of pure glycinin from soybeans

A novel method for the purification of glycinin from soy meal is presented. The method is based on the isoelectric precipitation of glycinin by using carbon dioxide as a volatile precipitant. Gaseous CO(2) was pressurized into the protein solution, thus lowering the pH and initiating glycinin precipitation. Pressurization and, consequently, acidification were done in a slow and controlled manner, with the end point of pH 6.4. The acidity of the protein solution was well controlled via the pressure of gaseous CO(2). In this way simultaneous precipitation of other soybean proteins was prevented and very pure glycinin was obtained. Approximately 40% of the glycinin present in the protein solution was recovered with purity as high as 98%. The purification process was successfully performed on both small and large scales, without affecting glycinin purity.     

Footrot and interdigital dermatitis in sheep: farmers' practices, opinions and attitudes

In 1999, a study was initiated to improve the treatment and control of footrot and interdigital dermatitis in sheep flocks in England and Wales. In November 2000, a retrospective cross-sectional study was conducted in which 392 sheep farmers were asked to estimate the prevalence of footrot and interdigital dermatitis in their flock in the previous 12 months, whether they considered these diseases to be a problem, how they treated and controlled them and their opinion on the success of the treatment and control measures that they used; 209 of them provided usable responses. The farmers tended to be more concerned as the prevalence of the diseases increased; 91 per cent of the farmers with a prevalence of footrot of less than 5 per cent considered it a small or very small problem on their farm, but 51 per cent of the farmers with a prevalence of 5 per cent or more also considered it to be a small or very small problem. Approximately 60 per cent of the farmers who used parenteral antibiotics considered that they were good or excellent at treating footrot, and this treatment was associated with a prevalence of less than 5 per cent. A similar proportion of farmers also considered topical foot sprays and footbathing to be good or excellent for controlling footrot or interdigital dermatitis, but these treatments were not associated with a lower prevalence of footrot or interdigital dermatitis. Of the 29 farmers who used a footrot vaccine, 20 (69 per cent) considered it good to excellent and this was associated with a prevalence of footrot of less than 5 per cent in their flock; however, vaccination was not associated with lower levels of footrot across the whole sample. Farmers spent approximately 34 minutes per sheep per year treating and controlling footrot; 31 per cent were prepared to spend more money and more time to manage footrot, 27 per cent were prepared to spend more time, 19 per cent were prepared to spend more money and 23 per cent were not prepared to do either. The farmers who were willing to invest more money or more money and time had a higher prevalence of footrot.     

Sustainability of a Privatized Community-based Animal Health Worker System in Mwingi District,Kenya

This paper describes a study on the sustainability of Community-based Animal Health Worker (CAHW) services in Mwingi District, Kenya. These services began in 1992 and were supported by the District Veterinary Authority (DVA) with assistance from the Integrated Food Security Programme – Eastern (IFSP-E). Over time and using a process of participatory reviews with multiple stakeholders, the system evolved into a network of CAHWs. The study focused on CAHWs service sustainability and their relationships with other animal health service providers. A mutually beneficial and supportive arrangement existed between the CAHWs and Animal Health Assistants (AHAs), based on a private drug supply system, referral and backstopping support. The CAHWs derived sufficient income from their veterinary work to maintain their interest in the system. Seventy per cent of CAHWs were continuing to offer adequate animal health services 3 years or more after their initial training and the withdrawal of donor support. Ninety-five per cent of sampled CAHWs (n = 40) viewed their business as successful and expanding. Considering the agro-ecological and socio-economic conditions of the district, the CAHW system can be viewed as an initial stage in the process of extending quality private sector veterinary services.     

Assessment of the Technical Competence and Ethical Behaviour of Community-based Animal Health Workers in Mwingi District,Kenya

This paper describes an assessment of the technical competence and ethical behaviour of Community-based Animal Health Workers (CAHWs) in Mwingi District, Kenya. From 99 trained CAHWs, 40 participated in the study. Using a pre-tested semi-structured questionnaire, direct observation of the relevant veterinary drug kits and participatory discussions, the study team found that the CAHWs knowledge of clinical signs of local livestock diseases and notifiable and zoonotic diseases and their ability to use veterinary drugs correctly and safely were adequate. Marks were awarded to the candidates according to an agreed marking scheme between the CAHWs trainers and study team members. The results showed that, overall, 36 out of 40 (90%) of the sampled CAHWs passed the tests. The existence of a referral system for CAHWs and refresher trainings helped to ensure that CAHW competence and ethical behaviour were maintained. However, it was also found that some areas of the current curriculum required more detailed input based on field experience. The CAHW system could serve as an alternative animal health care system in areas lacking veterinary services.     

A method to establish co-cultures of myotubes and preadipocytes from collagenase digested neonatal pig semitendinosus muscles

The relationships between adipocyte and muscle cell development within muscle are important in the study of factors or agents that may improve meat quality. Neonatal porcine muscle has the potential to yield both cell types for cell culture because it contains developing adipocytes and a high number of muscle satellite cells. Therefore, we modified a conventional collagenase-based procedure to digest neonatal porcine muscle and subsequently cultured the resultant muscle stromal-vascular (SV) cells on several substrata in basal and dexamethasone (DEX)-containing media. Developing myotubes and preadipocytes were present in muscle SV cell cultures on laminin substrata following seeding and plating with fetal bovine serum (FBS) with or without DEX. Myotube number was much higher (P < 0.05) on laminin substrata compared with all other substrata, whereas preadipocyte number in muscle SV cell cultures was independent of substrata, as we have shown previously. This approach can be used to establish co-cultures of differentiating adipocytes and myotubes from collagenase-digested neonatal pig muscle. Because the comparison is within the same culture dish, this method allows for a direct comparison of the responses of adipogenic and myogenic cells to growth and differentiation factors. For example, DEX did not alter myogenesis (i.e., 11 +/- 3 vs. 11 +/- 4 myotubes per unit area for control and DEX-treated cultures, respectively), but it has been shown to markedly increase preadipocyte number in muscle SV cell cultures.