Viral respiratory disease of the horse.

The diagnosis of any viral respiratory disease relies on laboratory procedures to isolate the virus and demonstrate a significant rise in serum antibody titers. To isolate viruses from the upper respiratory tract, it is imperative that nasopharyngeal swabs are obtained from animals in the early acute stage of illness, i.e., during the pyrexic phase when the virus is replicating. Nasopharyngeal swabs must be placed in a virus transport medium and forwarded immediately to the laboratory at refrigerated temperature. Equine influenza, rhinopneumonitis, and equine viral arteritis are the three viral infections causing outbreaks of respiratory disease in North America. African horse sickness, although foreign to North America, could be introduced despite stringent horse importation regulations. Specific antiviral therapy is not available to treat viral respiratory disease in the horse. A variety of inactivated and modified live vaccines, however, are available to prevent clinical disease and the spread of infection caused by the common viral respiratory pathogens. A considerable amount of research is underway to enhance the potency and duration of immunity of the present vaccines against influenza and rhinopneumonitis. This research is directed at defining and characterizing the importance of specific glycoprotein antigens on the surface of the virus, which trigger the various host immune responses, and determining whether they are stimulatory or suppressive.     

Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.     

Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Theileria orientalis: a review

Theileria orientalis (also known historically as T. sergenti and T. buffeli) is responsible for benign or non-transforming theileriosis, and exerts its major effect through erythrocyte destruction. The life cycle of T. orientalis is essentially similar to that of other Theileria species, except that the schizonts do not induce transformation and fatal lymphoproliferation. The pathogenesis of anaemia as a result of infection is not clearly established and may be multifaceted. Clinical signs of weakness, reluctance to walk and abortion are early but non-specific indications of disease, particularly if accompanied by a history of cattle being moved. Physical examination may reveal pallor (pale eyes, vaginal mucosa), pyrexia, and elevated heart and respiratory rates. T. orientalis is an economically important parasite of cattle in New Zealand, Australia and Japan, especially where naïve animals are introduced into an endemic area or in animals under stress. Increased awareness of the risks posed by the parasite is required to enable management practices to be implemented to minimise its impact.     

Prevalence of feline herpesvirus 1, Chlamydophila felis, and Mycoplasma spp DNA in conjunctival cells collected from cats with and without conjunctivitis

OBJECTIVE: To use PCR assays to determine the prevalence of feline herpesvirus 1 (FHV-1), Chlamydophila felis, and Mycoplasma spp DNA in conjunctival cells collected from cats with and without conjunctivitis; to compare results of conventional and real-time fluorogenic PCR assays for amplification of FHV-1 DNA; and to determine whether copy numbers of FHV-1 DNA are correlated with conjunctivitis. ANIMALS: 55 cats with active conjunctivitis, 39 healthy cats that never had conjunctivitis, and 32 cats with a history of conjunctivitis that had been resolved for at least 3 months. PROCEDURES: Samples were obtained by rolling cotton-tipped applicators on the ventral conjunctiva of awake cats treated topically with proparacaine. The DNA was extracted from the swab specimens and assessed in PCR assays to detect DNA of FHV-1 (fluorogenic PCR assay and conventional PCR assay), Mycoplasma spp (conventional PCR assay), and C felis (conventional PCR assay). RESULTS: Overall prevalence rates of FHV-1, C felis, and Mycoplasma spp as assessed by the conventional PCR assays were 6.7%, 3.2%, and 9.6%, respectively. Percentage concordance between conventional PCR and fluorogenic PCR assays for FHV-1 was 92.5%. There were no significant differences among the 3 groups of cats for the mean copy number of FHV-1 divided by the copy number of glyceraldehyde-3-phosphate dehydrogenase. CONCLUSIONS AND CLINICAL RELEVANCE: Mycoplasma spp were the most prevalent organism detected and was associated with conjunctivitis. This study could not confirm that there are increased copy numbers of FHV-1 DNA in cats with conjunctivitis, compared with the copy numbers for cats without conjunctivitis.     

Assessment of EST- and genomic microsatellite markers for variety discrimination and genetic diversity studies in wheat

It is likely that in the near future sequence information from sequencing programmes and EST libraries will generate an abundance of genic microsatellite markers. This study is focused on the assessment of their likely impact and performance vis-à-vis their genomic counterparts. Microsatellites from two sources were used to assess the genetic diversity in 56 old and new varieties of bread wheat on the UK Recommended List. A set of 12 microsatellite markers generated from genomic libraries and 20 expressed sequence tag (EST)-derived microsatellites were used in the study, and the performance of both marker sets assessed. The EST-derived or genic microsatellites delivered fingerprints of superior quality, amplifying clear products with few stutter bands. Diversity levels as revealed bygenic microsatellites are similar to the few published results. The PIC values for the genic markers were generally lower than those calculated for the genomic microsatellites, though advantages of both marker classes for variety identification applications are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Influence of food quality and quantity on the growth and development of Crassostrea gigas larvae: a modeling approach

A biochemically based model was developed to simulate the growth, development and metamorphosis of larvae of the Pacific oyster, Crassostrea gigas. The model is unique in that it (1) defines larvae in terms of their protein, neutral lipid, polar lipid, carbohydrate and ash content; (2) tracks weight separately from length to follow larval condition index and (3) includes genetic variation in growth efficiency and egg quality to better simulate cohort population dynamics. The model includes parameterizations for larval filtration, ingestion and respiration that determine growth rate and processes controlling larval mortality and metamorphosis. Changes in tissue composition occur as the larva grows and in response to the biochemical composition of the food.

The simulations show that genetically determined variations in growth efficiency produce significant changes in larval survival and success at metamorphosis. Larvae with low growth efficiency are successful under a much narrower range of culture conditions than larvae with high growth efficiency. The impact of low growth efficiency is primarily controlled by the ability of larvae to store lipid for metamorphosis. Culture conditions that provide increased dietary lipid counterweigh low growth efficiency. Changes in food quantity and quality had little effect on size at metamorphosis. On the other hand, larval life span and success rate at metamorphosis varied over a wide range depending upon the conditions of the simulation. Food quality and food availability both influence larval life span and, hence, larval survival. As ingestion rate decreases, larval life span increases and cohort survival declines. Increased lipid or decreased protein in the diet improves cohort survival. Changes in carbohydrate content are less influential. If cohort success is significantly affected by mortality during larval life rather than success at metamorphosis, the influence of food quality becomes more complex. The range of food compositions yielding high survival is restricted by a balance between improved success at metamorphosis obtained by increased lipid storage and the shortening of larval life span as a result of more rapid growth, a function of protein availability. These simulations illustrate the strength and utility of numerical models for evaluating and designing hatchery protocols for optimizing yield of C. gigas larvae.     

Salmon testes meal as a functional feed additive in fish meal and plant protein‐based diets for rainbow trout (Oncorhynchus mykiss Walbaum) and Nile tilapia (Oreochromis niloticus L.) fry

Fishery processing by‐products are a large resource from which to produce fishmeal and other products for a variety of uses. In this study, testes meal (TM) produced from pink salmon processing by‐product was evaluated as a functional ingredient in aquafeeds. Nile tilapia and rainbow trout fry were fed five isonitrogenous and isoenergetic experimental diets for 4 and 9 weeks respectively. Two diets were fishmeal‐based (FM) and three were plant protein‐based (PP). Salmon TM was added to the FM and PP diets at 7% to replace 20% of fishmeal protein (FMTM and PPTM respectively). An additional control diet was prepared in which fishmeal was added to the PP diet to supply an equivalent amount of protein as supplied by TM (PPFM). Inclusion of TM in both the FM‐ and PP‐based diets resulted in higher final body weights, although differences were only significant between rainbow trout fed FM or FMTM diets. Similar differences were calculated for other indices of fish performance, e.g. specific growth rate, feed conversion ratio, protein efficiency ratio and protein retention efficiency. Feed intake was significantly higher for fish fed FMTM compared with FM in rainbow trout. For tilapia, final weights were numerically higher, but not significantly different for fish fed diets containing TM compared with non‐TM diets (FM vs. FMTM; PP vs. PPTM). Performance of trout or tilapia fed the PPFM diet did not increase compared with the PP diet. The results indicate that TM addition to both FM and PP diets increased feed intake and also increased metabolic efficiency, demonstrating that TM can be a functional ingredient in aquafeeds.