Inter-laboratory and inter-assay comparison on two real-time PCR techniques for quantification of PCV2 nucleic acid extracted from field samples

Several real-time PCR assays for quantification of PCV2 DNA (qPCR) have been described in the literature, and different in-house assays are being used by laboratories around the world. A general threshold of 10(7) copies of PCV2 per millilitre serum for postweaning multisystemic wasting syndrome (PMWS) diagnosis has been suggested. However, neither inter-laboratory nor inter-assay comparisons have been published so far. In the present study, two different qPCR probe assays used routinely in two laboratories were compared on DNA extracted from serum, nasal and rectal swabs. Results showed a significant linear association between the assays (p<0.0001), and a systematic difference of 1.4 log10 copies of PCV2 per millilitre of sample (p<0.0001). This difference indicated that the assay from laboratory 1 yielded a higher output than the one from laboratory 2. Results also showed that there was no linear association between the amount of PCV2 DNA and the amount of total DNA, neither in nasal (p=0.86) nor in rectal (p=0.78) swabs, suggesting that normalizing of PCV2 DNA load in swab samples to total DNA concentration is not suitable. The present exploratory study highlights the need for the performance of ring trials on qPCV2 protocols between laboratories. Meanwhile, the proposed thresholds for PMWS diagnosis should only be considered reliable for each particular laboratory and each particular assay.     

The limitations of a feed/water based heat-stable vaccine delivery system forNnewcastle disease-control strategies for backyard poultry flocks in sub-Saharan Africa

Backyard poultry are a major contributor to egg and meat consumption in sub-Saharan Africa and an important source of income for many rural producers. Production throughout Africa is severely constrained by continuing outbreaks of Newcastle disease. The livestock-service sector lacks the resources and infrastructure to control Newcastle disease in extensive flocks without the active participation of producers. The development of 'heat-stable' Newcastle disease vaccines offers a potential solution. Trials over the last two decades have examined the effectiveness of heat-stable vaccines in both controlling Newcastle disease and in involving the rural community in control strategies. Constraints highlighted include the reliability of the vaccines using alternative delivery methods and the capacity of rural communities to apply those methods. The search for appropriate Newcastle disease-control strategies in extensive poultry systems should focus on policies and methodologies that incorporate the wider concerns and priorities of extensive producers.     

Flystrike prevention on Merino lambs with the insect growth regulator dicyclanil

The efficacy of a ready-to-use 5% dicyclanil pour-on formulation as a preventative of natural flystrike on Merino lambs was investigated in a field trial involving 5 sites in the southern Cape Province. A total of 1,804 lambs treated with dicyclanil were compared with 882 untreated control lambs for up to 25 weeks after treatment. Efficacy was assessed by calculating the weekly cumulative strike rate at each site and aggregated for all sites, as well as by calculating the percentage reduction achieved in treated lambs. Cumulative strike rates for the untreated controls aggregated for all sites at Weeks 9,14 and 19 after treatment reached 6.2, 12.8 and 17.8%, respectively, compared with 0.4, 1.7 and 3.6% for the dicyclanil treated lambs. Percentage control aggregated for all sites at Weeks 9,14 and 19 after treatment was 93, 87 and 80%, respectively. Heavy rains during the 1st half of the trial did not substantially reduce the efficacy of the product.     

Intravenous administration of levothyroxine for treatment of suspected myxedema coma complicated by severe hypothermia in a dog

A 7-year-old male English Coonhound with suspected myxedema coma complicated by severe hypothermia and metabolic abnormalities was treated with a combination of active external and core rewarming techniques, i.v. and oral administration of levothyroxine, supplemental oxygen, and administration of fluids (0.9% NaCl solution). Myxedema coma develops as a consequence of severe hypothyroidism and is characterized by a hypometabolic, stuporous state. Myxedema coma is associated with a high mortality rate, and most reported cases have involved Doberman Pinschers. Intravenous administration of levothyroxine can be used successfully in combination with oral administration to restore normal metabolic function and assist in warming and thermoregulation, although dosages should be conservative to avoid adverse cardiovascular effects.     

Ki-67 and minichromosome maintenance-7 (MCM7) expression in canine pituitary corticotroph adenomas

Pituitary-dependent hyperadrenocorticism (PDH) caused by pituitary corticotroph adenoma is a common endocrine disorder in dogs. The ratio between pituitary height and the area of the brain (P/B) has been used to evaluate the pituitary size. A P/B ratio > 0.31 indicates an enlarged pituitary, whereas a P/B ratio ≤ 0.31 indicates a nonenlarged pituitary. The aim of this study was to investigate the expression of proliferation markers Ki-67 and minichromosome maintenance-7 (MCM7) in canine corticotroph adenomas in enlarged and in nonenlarged pituitaries and to evaluate their relation with the size of canine pituitary corticotroph adenomas. Ki-67 and MCM7 expression in ACTH-positive tumor cells was determined by dual-labeling immunohistochemistry in resected corticotroph adenomas from 15 dogs with PDH. The mean ± SD Ki-67 labeling index (LI) was 0.55% ± 0.59% in corticotroph adenomas with nonenlarged pituitaries and 1.6% ± 0.6% in adenomas with enlarged pituitaries. The MCM7 LI in corticotroph adenomas with nonenlarged pituitaries and in adenomas with enlarged pituitaries was 2.9% ± 2.2% and 10.9% ± 3.7%, respectively. The Ki-67 LI and MCM7 LI were significantly greater in the adenomas with enlarged pituitaries than in the adenomas with nonenlarged pituitaries (P < 0.01 and P < 0.01, respectively). The MCM7 LI was significantly greater than the Ki-67 LI in adenomas (P < 0.01). The Ki-67 LI was positively correlated with the MCM7 LI (r = 0.820, P < 0.01), and the P/B ratio was positively correlated with the Ki-67 LI (r = 0.560, P = 0.03) and the MCM7 LI (r = 0.854, P < 0.01). In conclusion, canine corticotroph adenomas in enlarged pituitaries show greater proliferation potential than do adenomas in nonenlarged pituitaries. MCM7 expression was significantly greater than Ki-67 expression in canine pituitary corticotroph adenomas. Thus, MCM7 may be superior to Ki-67 as a proliferation marker in pituitary tumors.     

Influence of condensed tannin on intake, digestibility, and efficiency of protein utilization in beef steers fed high concentrate diet

This trial was conducted to evaluate if the effect of condensed tannin (CT) is associated with a true protein source on intake, ruminal and total digestibility, ruminal digestion rate, protein efficiency, and microbial efficiency in beef steers fed high concentrate diet (87% of DM). Four Bos indicus steers (407 ± 12 kg of BW) fitted with rumen cannula were assigned to a 4 × 4 Latin square design, arranged in a 2 × 2 factorial arrangement. Treatments consisted of either inclusion (0.4% of DM) or exclusion of condensed tannin (CT) from quebracho extract (76% of CT) with or without the use of soybean meal (SBM) as source of true protein. The level of inclusion was calculated to provide a daily intake of 1 g/10 kg of BW of CT. Intake of DM and nutrients was not affected (P > 0.10) by CT inclusion. However, there was an effect (P < 0.10) of CT inclusion on ether extract digestibility. An interaction (P < 0.10) was observed between CT and SBM on ruminal digestibility and digestion rate of crude protein (CP): when mixed with soybean meal, CT decreased the ruminal digestibility and, consequently, reduced the digestion rate of CP. Intake of CP increased (P < 0.10) with the inclusion of SBM. No differences in DM passage rate were observed (P > 0.10) among treatments. Effects of the interaction (P < 0.10) between CT and SBM were observed on flux of rumen undegradable protein (RUP), metabolizable protein (MP), and on the ratio MP:CP. In the presence of soybean meal, the addition of CT increased (P < 0.10) the flux of RUP and MP, and improved the ratio MP:CP. The yield of microbial protein on the abomasum and the microbial efficiency did not differ among treatments (P > 0.10). There was no difference (P > 0.10) on the pH, VFA, and ruminal ammonia (N-NH₃) with the addition of condensed tannin. The N-NH₃ increased and the ruminal pH decreased with the inclusion of soybean meal (P < 0.10). The utilization of condensed tannin as an additive in beef cattle diets with high level of concentrate and soybean meal as a source of true protein implies positive effects on crude protein utilization, decreasing digestion rate and ruminal digestibility of crude protein and consequently increasing the levels of metabolizable protein, with no changes in the ruminal fermentation parameters.     

Cutaneous and systemic blastomycosis, hypercalcemia, and excess synthesis of calcitriol in a domestic shorthair cat

A 9 yr old domestic shorthair cat was diagnosed with cutaneous and pulmonic blastomycosis. Severe persistent ionized hypercalcemia and excess circulating concentration of calcitriol were documented in association with blastomycosis. Ionized hypercalcemia resolved when the granulomatous lesions of blastomycosis resolved and the calcitriol levels decreased.     

Intersex condition of shoal bass in the Flint River, Georgia

We examined male shoal bass Micropterus cataractae from the Flint River, Georgia, to determine the prevalence of intersex. During March and April 2010, we sampled 61 shoal bass from six sites along the Flint River. Testes were examined histologically and classified as intersex if the presence of oocytes was noted. Using a severity index, we compared samples collected on different dates and from different locations according to age and testis weight. No significant variations were noted among any of the groupings. Further investigation is needed to determine whether the intersex condition in shoal bass is severe enough to warrant concern and whether it is a natural phenomenon.     

Radiosensitivity and capacity for radiation-induced sublethal damage repair of canine transitional cell carcinoma (TCC) cell lines

Understanding the inherent radiosensitivity and repair capacity of canine transitional cell carcinoma (TCC) can aid in optimizing radiation protocols to treat this disease. The objective of this study was to evaluate the parameters surviving fraction at 2 Gy (SF(2) ), α/β ratio and capacity for sublethal damage repair (SLDR) in response to radiation. Dose-response and split-dose studies were performed using the clonogenic assay. The mean SF(2) for three established TCC cell lines was high at 0.61. All the three cell lines exhibited a low to moderate α/β ratio, with the mean being 3.27. Two cell lines exhibited statistically increased survival at 4 and 24 h in the dose-response assay. Overall, our results indicate that the cell lines are moderately radioresistant, have a high repair capacity and behave similarly to a late-responding normal tissue. These findings indicate that the radiation protocols utilizing higher doses with less fractionation may be more effective for treating TCC.