A formal method of determining the dietary amino acid requirements of laying‐type pullets during their growing period

1. The amino acid requirements of laying type pullets during the growing period can be estimated by measuring the growth of different components of the body and making use of nutritional constants that define the amount of each amino acid that is required for the production of the tissues being formed.

2. In this experiment, carcase analyses of each of three breeds of pullets were conducted at weekly intervals throughout the growth of the pullets, to 18 weeks of age. Measurements were made of body weight, gut‐fill and feather weight, and chemical analyses consisted of water, protein, lipid and ash measurements of both the body and the feathers. Each age group comprised 10 birds of each breed.

3. Gompertz functions accurately estimated the growth of both body protein and feather protein, to 18 weeks of age, from which the rate of growth of these two components of the body could be estimated. The mature weight of pullets was overestimated by the Gompertz growth curve, which may indicate that a pullet ceases to increase in body protein content once sexual maturity has been reached.

4. Using allometric relationships between the chemical components of the body and of feathers, all the components of growth could be estimated from the growth of body protein and feather protein. These components were then added together to determine the growth rate of the body as a whole.

5. The daily amino acid requirements for 4 functions were calculated, namely, those for the maintenance of body protein and feather protein, and for the gain in body protein and feather protein. These requirements were then summed to determine the requirement of pullets on each day of the growing period.

6. Using the ‘effective energy’ system, the amount of energy required by these pullets was calculated for each day of the growing period, from which the desired daily food intake of the pullets could be predicted. By dividing the amino acid requirement by this daily food intake it was possible to determine the concentration of amino acids that would be needed in the diet in order to meet the requirements of a pullet.

7. The results indicate that the ratio between the requirement for lysine and for methionine and cysteine changes dramatically during the growing period, negating the concept of a fixed ratio between all the amino acids during growth.

8. The above process is the first step in determining the optimal feeding programme for a population of pullets of a given genotype. The constraining effects, of the diet being offered and of the environment in which the pullets are housed, on the food intake and growth rate of each pullet have to be estimated, and such a theory can then be expanded to include all the individuals in the population. Only by the use: of simulation models can all these constraining effects be considered simultaneously.     

Detection of Echinococcus multilocularis infection in a colony of cynomolgus monkeys (Macaca fascicularis) using serology and ultrasonography.

Five animals in a colony of cynomolgus monkeys (Macaca fascicularis) died or were euthanatized because of alveolar echinococcosis, during a period of 5 years. The remainder of the colony was screened for possible infection with Echinococcus multilocularis, using serology and ultrasonography. A total of 46 animals out of a group of 55 were examined. The presence of anti-Em2 antibodies analyzed with enzyme-linked immunosorbent assay was demonstrated in 3 monkeys. In 2 of these 3 monkeys, multilocular structures compatible with metacestodal cysts in the liver were identified, using ultrasonography. The presence of alveolar echinococcosis was subsequently confirmed at postmortem examination in 1 animal. The other animals are still alive. Two other monkeys were negative in the serological examination but had cystic structures in the liver, which were identified as bile duct cysts at postmortem examination in 1 animal. The other monkey is still alive. These findings suggest that serology for antibodies against the Em2 antigen may represent a useful method in identifying animals that might be infected with E. multilocularis and are therefore at risk of developing fatal alveolar echinococcosis.     

Non-structural plum pox potyvirus proteins detected by immunogold labelling

Plum pox potyvirus (PPV) induces in infected Nicotiana clevelandii cells characteristic crystalline inclusions known as nuclear inclusions (NI) when located in the nucleus and as dense material (Dm) when located in the cytoplasm. Crystalline inclusions contain protease (NIa) and RNA-dependent RNA polymerase (NIb) proteins. It is now well established for all potyviruses that cylindrical inclusions contain CI helicase ATPase protein (Martin et al., 1992). The intracellular location of other non-structural PPV proteins remains unknown. Using Escherichia coli expression vectors, specific antibodies were obtained against P1, P3, 6K2 and NIb PPV proteins for which antibodies were not yet available. As expected, NIb antiserum labelled crystalline inclusions. P1, P3 and 6K2 proteins were present in both types of crystalline inclusions found in the nucleus and in the cytoplasm of PPV-infected leaves of N. clevelandii, suggesting that nuclear inclusions and dense material were composed of the same proteins. This composition is discussed.     

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.     

Immunohistochemical differentiation of reactive from malignant mesothelium as a diagnostic aid in canine pericardial disease

Objectives

To develop a provisional immunohistochemistry panel for distinguishing reactive pericardium, atypical mesothelial proliferation and mesothelioma in dogs.

Materials and Methods

Archived pericardial biopsies were subject to haematoxylin and eosin staining, immunohistochemistry for cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Samples were scored for intensity and number of cells stained.

Results

Ten biopsies of reactive mesothelium, 17 of atypical mesothelial proliferation, 26 of mesothelioma and five of normal pericardium were identified on the basis of haematoxylin and eosin staining. Cytokeratin and vimentin were expressed in all biopsies, confirming mesothelial origin. Normal pericardial samples had the lowest scores for insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Mesothelioma and atypical proliferative samples were similar to each other, with higher scores for insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 than the reactive samples. Desmin staining was variable. Insulin‐like growth factor II mRNA‐binding protein 3 was the best to distinguish between disease groups.

Clinical Significance

An immunohistochemistry panel of cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 could provide superior information compared with haematoxylin and eosin staining alone in the diagnosis of cases of mesothelial proliferation in canine pericardium, but further validation is warranted.