Sexing chick embryos: a rapid and simple protocol

1. Analysis of gene expression in the developing chick gonads requires the collection of male and female tissues from embryos between 3.5 d and 8.5 d of development. However, male and female chick embryos are indistinguishable by morphological examination before d 7.5 of development. 2. Sex identification of earlier embryos is only possible by molecular methods, which at present are laborious and time consuming. 3. We have devised a PCR-based sexing protocol which combines both sex specific and control reactions in a single tube assay. The assay is rapid and effective over a wide range of DNA concentrations and is tolerant of poor quality DNA. 4. Procedures are described for identifying the sex of individual embryos using either tissue samples or a small number of cells recovered from amniotic fluid.     

Immune response of chickens inoculated with a recombinant avian leukosis virus

Specific-pathogen-free embryos (18-day incubation) and hatched chicks were inoculated with a recombinant avian leukosis virus (ALV) produced by recombinant DNA techniques. Enzyme-linked immunosorbent assays were used to measure the production of viral-protein-specific antibody and the viral protein, p27, in the serum at 2, 5, 8, 14, and 20 weeks of age. Of the inoculated chickens surviving to 20 weeks, 64% produced viral-protein-specific antibodies and 42% transiently produced the viral protein, p27. Chickens inoculated as embryos did not differ significantly from those inoculated at hatch with respect to antibody and viral protein production. Antibody production peaked at 5 weeks postinoculation and declined over the remaining 15 weeks of the study. No evidence of chronic tolerant infection or mortality due to neoplastic disease was found.     

Enzyme studies with the livers of chicks fed semi‐synthetic diets containing crystalline amino acids and diammonium citrate

The levels of glutamate dehydrogenase [NAD(P)], (GDH), aspartate trans‐aminase (AspT) and alanine transaminase (AlT) were measured in livers from chicks fed on a semi‐synthetic diet containing crystalline essential amino acids as the sole nitrogen source (diet A). The effects of a supplement of 12.0% glutamic acid (diet H) or 11.07% diammonium citrate (DAC) (diet B) or 12.0% glutamic acid plus 1.0% proline with an additional 0.6% glycine (diet C) on these enzymes were studied and the results compared with the levels found for control chicks given a typical diet based on cereal protein (diet J). The abilities of livers from chicks given diets A, B and C to synthesise [14C] glutamic acid from [14C]2‐oxoglutaric acid and diammonium citrate (DAC) were assessed.

The levels of GDH, AspT and AlT found in the livers of chicks given the control diet were 54.1, 966 and 123.7 units/mg protein respectively. Non‐essential nitrogen added as glutamic acid or as DAC did not cause induction of the enzymes studied above control levels. Glutamic acid (diets H and B) caused a depression of GDH levels (37.4 and 38.9 units/mg protein respectively) but had no effect on AspT and AlT compared with the controls, whereas DAC caused a decrease in AlT (43.9 units/mg protein) but had no effect on AspT and GDH; diet A depressed AlT and AspT levels (64.4 and 735 units/mg protein respectively).

The livers of chicks given diets A, C and B varied in their ability to synthesise glutamic acid, 39.3%, 31.9% and 24.0% respectively of the radioactivity being recovered as glutamic acid.     

Effect of oral administration of electrolyte pastes on rehydration of horses

OBJECTIVE: To determine whether the composition of electrolyte pastes formulated for oral administration influences voluntary water intake (WI) by horses recovering from furosemide-induced dehydration. ANIMALS: 6 horses. PROCEDURES: Voluntary WI, body weight, and blood and urine constituents were measured before and after induction of dehydration by furosemide administration and overnight withholding of water; these same variables also were measured during a 36-hour rehydration period. Each horse was evaluated 4 times with random application of 4 treatments (electrolyte pastes) that provided 0.5 g of KCl/kg of body weight, 0.5 g of NaCl/kg, 0.25 g of NaCl and 0.25 g of KCl/kg, or no electrolytes (control treatment). Electrolyte pastes were administered 3 times (4, 8, and 12 hours after start of the rehydration period). RESULTS: Administration of all electrolyte pastes resulted in significantly greater voluntarily WI, compared with the control treatment, and was accompanied by significantly greater recovery of body weight when NaCl was a component of the paste. Administration of NaCl and NaCl-KCl pastes tended to produce a state of transient hyperhydration; however, electrolyte administration also resulted in significantly greater urine production and electrolyte excretion during the final 24 hours of the rehydration period. Adverse effects of oral administration of hypertonic electrolyte pastes were not observed. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of electrolyte pastes to dehydrated horses increases voluntary WI and improves rehydration during the rehydration period. Rehydration is more rapid and complete when NaCl is a component of the electrolyte paste.     

Weaning,yearling, and preharvest ultrasound measures of fat and muscle area in steers,bulls, and heifers

Longissimus muscle area and fat thickness were measured following weaning, at yearling, and prior to harvest using real-time ultrasound, and corresponding carcass measurements were recorded 3 to 7 d following the preharvest scan in composite steers (n = 116, 447 +/- 19 d), bulls (n = 224, 521 +/- 11 d), and heifers (n = 257,532 +/- 12 d). Although fat deposition was limited in bulls and heifers from weaning to yearling, coefficients of variation ranged from 8.46 to 13.46% for muscle area, and from 27.55 to 38.95% for fat thickness, indicating that significant phenotypic variance exists across genders. Residual correlations, adjusted for the effects of year of birth, gender, and age at measurement, were high and ranged from 0.79 to 0.87 among ultrasound and carcass measures of muscle area. Residual correlations among ultrasound and carcass measures of fat thickness were also high, ranging from 0.64 to 0.86. Weaning and/or yearling ultrasound muscle area yielded similarly accurate predictions of carcass muscle area. Yearling ultrasound fat thickness accounted for 13% more of the observed variance in carcass fat thickness than the weaning ultrasound measure in single-trait prediction models. When both weaning and yearling ultrasound measures were used to predict carcass fat thickness, partial R2 values were 0.15 and 0.61 for weaning and yearling ultrasound fat thickness, respectively. The difference between predicted and carcass measures with respect to muscle area (fat thickness) was less than 6.45 cm2 (2.5 mm) for 80.2 to 88.9% (90.3 to 95%) of animals. Preharvest ultrasound measures yielded standard errors of prediction of less than 4.95 cm2 for muscle area and 1.51 mm or less for fat thickness. These results indicate that ultrasound measures taken between weaning and yearling provide accurate predictors of corresponding carcass traits in steers, bulls, and heifers.     

The Effects of Replacing Dicalcium Phosphate with Busumbu Rock Phosphate on Performance and the Mechanical Properties of Bone in Growing Chicks

Three hundred, day-old broiler chicks, with an average initial weight of 41.8±1.79 g, were used in a 15-day study (10 birds per battery cage) to characterize their performance and fluorine status when dicalcium phosphate (DCP) was replaced by Busumbu rock phosphate (BRP) as the source of phosphorus in the chicks' ration. The treatments comprised a standard ration with BRP replacing 0, 25%, 50%, 75% or 100% of DCP. Replacing DCP with BRP significantly reduced the final weight of the chicks (p<0.01), feed intake (p<0.01), weight gains (p<0.01) and dry matter digestibility (p<0.05) but increased (p<0.05) the feed-to-gain ratio. True phosphorus absorption and the percentage of phosphorus in the tibia were not affected by increasing amounts of BRP in the diet. Increasing levels of BRP in the diet linearly reduced (p<0.01) the percentage bone ash, calcium, Ca:P ratio, ultimate breaking force, bending moment, stress, and modulus of elasticity. Leg stiffness, lameness, reduced feed intake, and a decline in general health were recorded in 10–40% of the chicks on 75% and 100% BRP, respectively. These results suggest that excessive ingestion of fluorine from the BRP caused the reduction in the chicks' performance.     

Intralesional cisplatin chemotherapy and topical cryotherapy for the control of choanal squamous cell carcinoma in an African penguin (Spheniscus demersus).

A 3.5-yr-old, 2.5-kg female African penguin (Spheniscus demersus) was diagnosed with a choanal squamous cell carcinoma on the basis of biopsy after a history of choanal ulceration and anorexia with weight loss. Therapeutic modalities included the use of intralesional cisplatin followed by localized, topical cryotherapy with liquid nitrogen. The penguin remained free of clinical disease for a period of 9 mo, at which time a second cryotreatment was performed. The patient has remained free of all signs of choanal carcinoma for a period of 13 mo since the second cryotreatment.     

Toward a multiplexed serotyping immunoassay for foot-and-mouth disease virus.

Initial results demonstrating the feasibility of a multiplexed liquid array immunoassay for foot-and-mouth disease viral antigen detection and simultaneous serotype differentiation are presented. Serotype-specific antibodies from rabbit and guinea pig hyperimmunesera were isolated and prepared for use in a multiplexed, bead-based assay. The performance of all of the available antibodies as both capture and detector reagents was evaluated in the multiplexed system to establish a combination exhibiting the highest homotypic responses and lowest heterotypic reactions. The multiplexed assay was evaluated against inactivated cell culture supernatant samples of the same subtype as the virus used to raise the capture and detector antibodies. Distinct serotype differentiation was observed, except in the case of serotype SAT1. Subsequently, cell culture supernatant samples from a larger pool of viral subtypes were analyzed. Distinct serotype differentiation was obtained when analyzing cell culture supernatant samples from viral serotypes C, Asia, and SAT3, irrespective of the subtype. However, limitations of the current antibody pairs were realized in some inconclusive results obtained when analyzing samples from a broader range of O, A, and SAT2 subtypes. The results obtained in this initial study will be used to further optimize the assay using polyvalent or monoclonal antibodies and move toward the analysis of clinical samples.     

Growth and metabolism in somatotropin-treated steers: III. Protein synthesis and tissue energy expenditures

Rates of in vivo protein synthesis in intercostal, sartorius and semitendinosus muscle and in the heart, liver, kidneys, rumen and jejunum were determined in 20 growing Hereford steers treated for 112 d with daily subcutaneous injections of either saline (S) or recombinantly derived bovine somatotropin (rBST; 20.6 mg/d). In vitro rates of protein synthesis and energy expenditures associated with Na+, K+ transport also were determined in external intercostal muscle, liver, kidneys and jejunum. Neither in vivo nor in vitro tissue protein fractional synthetic rates (mg/[g protein.d]), using either the plasma (P) or intracellular fluid (ICF) phenylalanine specific radioactivity for the precursor pool, were affected by rBST treatments. Energy expended on Na+, K+ transport was greater (P less than .1) in the livers of rBST-treated steers, which would increase the maintenance energy expenditures of these steers. Protein accretion rates in the liver, kidneys, stomach, hide, and head, feet and tail of rBST-treated steers were greater (P less than .05) than in S steers. Tissue amino acid profiles were not affected by rBST treatments except in the rumen, where profiles suggested that less collagen was present in rumen wall tissue protein of rBST steers. Plasma phenylalanine entry rates also were not affected by rBST treatment; muscle protein synthesis accounted for a minimum of 20% of this entry rate.     

The effect of cattle foot traffic on soil compaction in a silvo-pastoral system

In India, small farmers have generally found that the growing of eucalypts as a cash crop provides poor returns, and many have reverted to growing annual crops. Farmgate prices have been much lower than retail prices, due to the existence of middlemen, and retail prices have fallen to lower levels than expected as markets have become saturated. This article describes the situation in eastern Gujarant, where farmers sold their eucalypts directly to buyers at prices close to those in organised wood markets. An important reason for this unusual situation is the absence in Gujarat of controls on eucalyptus sales, which has helped producers to avoid dependence on rent-seeking intermediaries. The survey shows that growing eucalyptus can be a profitable activity for small farmers, given the right circumstances.