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.     

Effects of estradiol 17 beta implants on hematologic values and the chemiluminescence response of neutrophils of steers.

The effects of subcutaneous administration of a commercially available estradiol 17 beta implant on hematologic values and the chemiluminescence response of neutrophils were evaluated in 14 steers. Chemiluminescence and hematologic values were measured in treated (n = 8) and nontreated (n = 6) steers on days -14, -7, and -1 prior to implantation. Estradiol 17B was implanted into the treated group of steers on day 0, and blood samples were obtained from all steers on days 1, 2, 3, 4, 8, 15, 22, 29, 36, 43, and 50. The concentration of estrogen in serum was significantly (P = 0.0120) higher following implantation. Chemiluminescence and hematologic indices were not significantly affected by either implant status or serum concentrations of estrogen. The results of this study suggested that the use of implants containing estradiol 17 beta for promotion of weight gain in steers will not result in alterations of hematologic values or the neutrophil respiratory burst.     

The control of Fannia canicularis in a poultry house using a black-light technique

The lesser house fly Fannia canicularis and other species of fly of animal health and public health importance, were shown to be attracted to “ black‐light “ traps in semi‐open poultry houses permitting free access and strong daylight competition. The fly populations were reduced and the number of ovipositions in the droppings pit apparently decreased. It is shown that the use of these traps, normally reserved for indoor insect control, can be extended to include certain farm situations.     

Mycobacterium infection in a gray-cheeked parakeet

A gray-cheeked parakeet, approximately 3 years of age, was submitted for necropsy. Clinical signs included diarrhea, decreased feed consumption, fluffing of feathers, shivering, weight loss, and limping. Necropsy revealed enlargement and pale discoloration of the spleen and intestines. The liver contained multifocal pale foci. Granulomas, some of which had necrotic centers, were present along the subserosa of the intestinal tract. Microscopic examination revealed diffuse granulomatous splenitis, enteritis, and multifocal granulomatous hepatitis. Numerous acid-fast bacteria were demonstrated in the spleen, liver, and intestine.     

Diagnosis of sporotrichosis in a donkey using direct fluorescein-labeled antibody testing.

A 4-year-old female donkey residing in an open field in Indiana was admitted for evaluation of facial lesions of 2 years duration. Cytologic and histologic examination of exudate and tissue from the lesions revealed a pyogranulomatous inflammatory reaction with numerous yeasts. Sporothrix schenckii was suspected to be the infectious agent; however, multiple culture attempts did not provide positive identification of the organism. Serologic examination supported infection with S. schenckii. A specific direct immunofluorescent antibody test performed on paraffin-embedded tissue sections confirmed the organism as S. schenckii. Clinical signs resolved after appropriate iodide therapy.     

Assessing economic and social pressure for the control of bovine viral diarrhoea virus

The objective of this paper is to present a preliminary assessment of variation in the economic impact of bovine viral diarrhoea virus (BVDV) at dairy farm level between a sample of nations within the EU and hence assess differences in pressure to respond to this disease that may be impeding progress in control and hence restricting collective benefits from healthier livestock. We used a questionnaire to obtain national average values of key epidemiological and economic parameters for a typical dairy farm from BVDV experts in the countries concerned. These parameters were converted into assessments of economic impact using a computer simulation model. Uncontrolled output losses for a BVDV-na?ve herd with virus introduced in year 1 of a 10-year epidemic represented 22, 7, 8, 5, 8 and 20% of the BVDV-free annuity for the UK, Northern Portugal, Holland, Norway, Italy and Germany, respectively. Differences between countries will be widened by differences in the risk of acquiring BVDV. These will be much reduced in countries, such as Norway that have a national BVDV eradication programme. Farmers in such countries can therefore justify spending much less on maintaining BVDV-free status than BVDV-free farms in other countries. This result illustrates the paradox that in countries where BVDV prevalence is high, farmers have least to gain from unilateral BVDV eradication because of the high cost of maintaining freedom from the disease. We discuss this issue in the light of increasing recognition at international level of the importance of BVDV control.     

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.