Detection of genomic DNA of the crayfish plague fungus Aphanomyces astaci (Oomycete) in clinical samples by PCR

A diagnostic procedure, based on a polymerase chain reaction method (PCR) was developed to detect infection of crayfish with the Oomycete Aphanomyces astaci. A set of oligonucleotide primers was designed to specifically amplify A. astaci DNA in the ITS region surrounding the 5.8S rDNA gene. The PCR amplifies a 115 bp amplicon. The specificity of the primers was demonstrated by testing on 27 A. astaci strains and against 20 non-A. astaci Oomycetes and 5 fungal species. Most of the non-A. astaci Oomycete or fungal species included in the study are either known parasites of freshwater crayfish cuticle or can be found in their natural environment. Specificity was also tested against crayfish tissue and some known parasites and bacteria infecting crayfish.

A protocol for the extraction of A. astaci DNA from infected crayfish tissue was developed. The optimised method allows the detection of two genome equivalents of purified A. astaci genomic DNA.

The method was tested on noble crayfish (Astacus astacus), artificially infected with A. astaci. Detection of A. astaci was possible at the very first time of sampling, which was 2 days after the beginning of spore exposure.     

A mechanistic model for predicting the nutrient requirements and feed biological values for sheep

The Cornell Net Carbohydrate and Protein System (CNCPS), a mechanistic model that predicts nutrient requirements and biological values of feeds for cattle, was modified for use with sheep. Published equations were added for predicting the energy and protein requirements of sheep, with a special emphasis on dairy sheep, whose specific needs are not considered by most sheep-feeding systems. The CNCPS for cattle equations that are used to predict the supply of nutrients from each feed were modified to include new solid and liquid ruminal passage rates for sheep, and revised equations were inserted to predict metabolic fecal N. Equations were added to predict fluxes in body energy and protein reserves from BW and condition score. When evaluated with data from seven published studies (19 treatments), for which the CNCPS for sheep predicted positive ruminal N balance, the CNCPS for sheep predicted OM digestibility, which is used to predict feed ME values, with no mean bias (1.1 g/100 g of OM; P > 0.10) and a low root mean squared prediction error (RMSPE; 3.6 g/100 g of OM). Crude protein digestibility, which is used to predict N excretion, was evaluated with eight published studies (23 treatments). The model predicted CP digestibility with no mean bias (-1.9 g/100 g of CP; P > 0.10) but with a large RMSPE (7.2 g/100 g of CP). Evaluation with a data set of published studies in which the CNCPS for sheep predicted negative ruminal N balance indicated that the model tended to underpredict OM digestibility (mean bias of -3.3 g/100 g of OM, P > 0.10; RMSPE = 6.5 g/100 g of OM; n = 12) and to overpredict CP digestibility (mean bias of 2.7 g/100 g of CP, P > 0.10; RMSPE = 12.8 g/100 g of CP; n = 7). The ability of the CNCPS for sheep to predict gains and losses in shrunk BW was evaluated using data from six studies with adult sheep (13 treatments with lactating ewes and 16 with dry ewes). It accurately predicted variations in shrunk BW when diets had positive N balance (mean bias of 5.8 g/d; P > 0.10; RMSPE of 30.0 g/d; n = 15), whereas it markedly overpredicted the variations in shrunk BW when ruminal balance was negative (mean bias of 53.4 g/d, P < 0.05; RMSPE = 84.1 g/d; n = 14). These evaluations indicated that the Cornell Net Carbohydrate and Protein System for Sheep can be used to predict energy and protein requirements, feed biological values, and BW gains and losses in adult sheep.     

Identification of a novel hemotropic mycoplasma in a splenectomized dog with hemic neoplasia

A 3-year-old sexually intact male Bull Mastiff underwent splenectomy for splenic thrombosis; prior to and after splenectomy, multiple blood transfusions were administered. Two weeks after the procedure, T-cell lymphoproliferative disease was diagnosed. Treatment with prednisone and chlorambucil was initiated, and 2 weeks later, cytologic examination of a blood smear revealed small (0.3 microm), coccoid basophilic bodies on the surface of approximately 70% of the RBCs. Morphologically, these resembled "Candidatus Mycoplasma haemominutum." A polymerase chain reaction assay was used to amplify a partial 16S rRNA sequence in blood obtained from the dog; the product was sequenced and compared with 16S rRNA gene sequences of other hemotropic mycoplasmas. The sequence was 98% homologous to that of "Candidatus M haemominutum", but only 77% homologous to that of M haemocanis and M haemofelis.     

Veterinary school admission interviews, part 3: strategies for increasing interview validity

The veterinary school admission interview is a widely used selection tool, yet concerns persist about its reliability, validity, and cost. Relative to medicine, optometry, and dentistry schools, veterinary schools have been more likely to conduct panel interviews and to fix the interview's weight in selection decisions, strategies that increase interview validity. This article provides strategies for further increasing the veterinary school interview's validity. Interview reliability and validity studies point to key strategies the veterinary school admissions committee can implement before the interview: (1) establishing the interview's purpose(s); (2) conducting a "job" analysis to identify desirable candidate skills, knowledge, and attributes; (3) developing a structured and panel interview where interviewers, if possible, are blind to other admission data; (4) training interviewers; (5) setting a reasonable interview schedule; and (6) determining methods for analyzing applicant data. During the interview, interviewers should proceed through a structured series of steps: (1) open the interview with a specified agenda; (2) probe for information using structured questions and anchored rating scales; (3) close the interview to allow for candidate questions; and (4) evaluate the interview data. After the interview, the admissions committee should (1) analyze the interview data within and across interviewers and (2) analyze the data across all selection tools in order to assign relative weights to the selection tools.     

Effects of contact dermatitis on hepatic gene expression in broilers

1.?Severe foot and hock dermatitis in broiler chickens can reduce growth rate and increase susceptibility to bacterial infection, affecting both profitability and welfare. However, little is known about the underlying physiological changes associated with foot and hock lesions.

2.?This study compared global hepatic gene expression in control birds and those with ammonia-induced foot and hock lesions using Agilent 44?K chicken oligonucleotide microarrays (8 birds per group).

3.?In total, 417 genes were differentially expressed of which 174 could be mapped onto the genome. Genes associated with energy metabolism, thyroid hormone activity and cellular control were affected, while there was also evidence of an up-regulation of genes linked to a pro-inflammatory response.

4.?It is conceivable that pain is the underlying cause for the observed changes in energy metabolism genes.

5.?Changes in hepatic gene expression provide new information on how a chicken's physiological mechanisms alter to cope with foot and hock lesions. The findings support other data indicating that birds with increased severity of lesions are likely to be in pain and that growth will be compromised. Reduction of the incidence of dermatitis by improved husbandry should therefore benefit both welfare and commercial performance.     

Sodium salicylate attenuates lipopolysaccharide (LPS)-induced adipsia, but not hypophagia, in broiler chickens

A study was conducted to determine the influence of sodium salicylate on the behaviour and the food and water consumption of broiler chickens after lipopolysaccharide (LPS) injection. An oral dose of 100 mg/kg sodium salicylate was given and an acute phase reaction in broiler chickens was provoked through the intravenous injection of Escherichia coli LPS. Water intake was higher in the LPS and salicylate-treated group than in the positive control group. The salicylate treatment, however, did not restore the food intake, or influence the behaviour of the chickens. These data show that sodium salicylate has a positive effect on the water intake after intravenous injection of LPS in chickens and suggests that there is a difference in mechanism of action of food and water consumption after LPS injection in chickens.     

Validation of the FAMACHA eye color chart for detecting clinical anemia in sheep and goats on farms in the southern United States

Recent studies on sheep and goat farms in the southern United States indicate that multiple-anthelmintic resistance in Haemonchus contortus is becoming a severe problem. Though many factors are involved in the evolution of resistance, the proportion of the parasite population under drug selection is believed to be the single most important factor influencing how rapidly resistance develops. Therefore, where prevention of resistance is an important parallel goal of worm control, it is recommended to leave a portion of the animals untreated. Recently, a novel system called FAMACHA was developed in South Africa, which enables clinical identification of anemic sheep and goats. When H. contortus is the primary parasitic pathogen, this system can be applied on the farm level to reduce the number of treatments administered, thereby increasing the proportion of the worm population in refugia. Since most studies validating the FAMACHA method have been performed in South Africa, it is important that the method be tested in other regions before its use is broadly recommended. We performed a validation study of FAMACHA by testing the system in sheep (n = 847) and goats (n = 537) of various breeds and ages from 39 farms located in Arkansas, Georgia, Louisiana, Florida, and the US Virgin Islands. The color of the ocular conjunctiva of all animals were scored on a 1-5 scale using the FAMACHA card, and blood samples were collected from each animal for determination of packed cell volume (PCV). Fecal samples were also collected from a majority of the animals tested for performance of fecal egg counts (FEC). Correlations between PCV and eye scores, PCV and FEC, and FEC and eye scores were all highly significant for both sheep and goats (P < 0.001). Data for both FAMACHA scores and PCV were evaluated using two separate criteria for anemia: eye score values of 3, 4 and 5 or 4 and 5, and PCV values of < or =19 or < or =15 were considered anemic. Specificity was maximized when eye score values of 4 and 5 were considered anemic and PCV cut off for anemia was < or =19, but sensitivity was low. In contrast, sensitivity was 100% for both sheep and goats when eye score values of 3, 4 and 5 were considered anemic and PCV cut off was < or =15, but specificity was low. In both sheep and goats, predictive value of a negative was greater than 92% for all anemia and eye score categories, and was greater than 99% for both eye score categories when an anemia cutoff of < or =15 was used. Predictive value of a positive test was low under all criteria indicating that many non-anemic animals would be treated using this system. However, compared to conventional dosing practices where all animals are treated, a large proportion of animals would still be left untreated. These data indicate that the FAMACHA method is an extremely useful tool for identifying anemic sheep and goats in the southern US and US Virgin Islands. However, further studies are required to determine optimal strategies for incorporating FAMACHA-based selective treatment protocols into integrated nematode control programs.