Technical note: mineral deposits on Dacron bags during ruminal incubation.

Extensive DM contamination was found on Dacron bags that were incubated for prolonged periods of time in the rumen of steers fed alfalfa hay. The ash content of the contaminant was high, and most of it was acid-soluble X-ray analysis indicated the presence of hydroxylapatite and synthetic calcium magnesium phosphate or whitlockite. The contaminant appeared as a smooth coating on the Dacron fiber, suggesting that contamination was a gradual process rather than the result of entrapment of dislodged crystals from plant material. Contamination seemed to occur exponentially within the range of observations (0 to 42 d). Contamination also occurred in steers fed orchardgrass, although to a lesser extent than in steers fed alfalfa hay. The DM contamination was less than .04 g per bag (average bag weight was 1.2 g) during the first 10 d of incubation. However, correction for contamination might be required for studies involving longterm incubation or mineral digestion.     

Manipulation of nitrogen digestion by sheep using defaunation and various nitrogen supplementation regimens

Five ruminally, duodenally, and ileally cannulated sheep (average BW 62 kg) were fed 65% roughage: 35% concentrate diets (CP = 15%) in a 5 x 5 Latin square design to study the applicability of using a combination of defaunation with N supplements (soybean meal [SBM], corn gluten meal [CGM], blood meal [BM], urea, and casein) with different extents of ruminal degradation to manipulate microbial protein synthesis and amount of ruminal escape protein. Diets were fed twice daily (1,759 g DM/d). Defaunation was accomplished with 30-ml doses of alkanate 3SL3 (active ingredient: sodium lauryl diethoxy sulfate)/sheep daily for 3 d with 2 d of fasting. Treatment 1 (control) involved feeding faunated sheep a diet in which the supplemental N (45% of total dietary N) was 67% SBM N and 33% urea N. Treatment 2 involved feeding defaunated sheep the same diet as the control. Treatments 3, 4, and 5 involved feeding defaunated sheep diets in which the supplemental N source was either 67% CGM-BM (1:1 N ratio) N:33% urea N, or 33% CGM-BM N:67% urea N or 33% CGM-BM N:33% urea N:33% casein N, respectively. Compared with the faunated control, defaunation decreased (P less than .05) ruminal ammonia concentration (19 vs 26 mg/dl) and increased (P less than .05) CP flow to the duodenum (253 vs 214 g/d) due to a trend for increases in both bacterial (BCP) and nonbacterial (NBCP) CP flows.(ABSTRACT TRUNCATED AT 250 WORDS)     

On Deep Freezing of Boar Semen: Investigations on the Effects of Different Straw Volumes, Methods of Freezing and Thawing Extenders

Contents: At the Bundesanstalt Wels tests with frozen-thawed boar semen in plastic straws were conducted. The influences of straw volume, method of freezing and thawing extender were investigated. The straw volumes of 1, 0 and 0, 5 ml showed significantly better thawing results in a preliminary trial than the 5, 0 ml Macrotub. When semen was frozen in a computerized freezer with automatic seeding, all tested straw. volumes gave significantly better results than the straws frozen in static N₂-vapor. A ready to use commercial extender was used for thawing with good results to simplify the handling of deep frozen semen for on farm insemination. In the main trial 60 ejaculates from 10 boars were frozen in 1, 0 ml straws in the computerized freezer. Three in vitro parameters for fresh semen (motility, osmotic resistance and keeping quality under standard conditions) and two parameters for thawed semen (motility and percentage of sperm with normal apical ridge) were recorded and correlated. The osmotic resistance test proved to be well suited as means of predicting the fitness of an ejaculate for deep freezing, but the other two fresh semen parameters showed poor correlation with the parameters for thawed semen. From the parameters for thawed semen a freezing score was derived as a measure of the freezability of single ejaculates and boars. A preliminary insemination trial gave satisfying farrowing rates. Inhalt: Zur Tiefgefrierung von Ebersamen: Untersuchungen zum Einfluß von verschiedenen “Straw”-Volumina, Einfriewerfahren und Auftaubedingungen Ander Bundesanstalt Wels wurden Tiefgefrierversuche mit Eberspermain Kunststoffpailletten durchgeführt. Die jeweiligen Einflüsse von Paillettenvolumen, Gefrierverfahren und Auftauverdünner wurden untersucht. In Vorversuchen erwiesen sich die Paillettenvolumina 1, 00 und 0, 5 ml gegenüber den 5,0 ml Makrotüb hinsichtlich Kopfkappenintegrität und Auftaumotilität signifkant überlegen. Die Auftauergebnisse der im computergesteuerten Freezer mit automatischem Seeding eingefrorenen Proben waren für alle untersuchten Paillettengröβen signifikant bis hoch signifikant besser, als die Werte der im statischen Stickstoffdampf eingefrorenen Proben. Als Auftauverdünner wurde ein handelsüblicher Fertigverdünner gewählt um eine besonders für den Eigenbestandsbesamer wichtige einfache Handhabung der TG-Besamung zu gewährleisten. Im Hauptversuch wurden 60 Ejakulate von 10 Ebern verwendet und in 1, 0 ml Pailletten im programmierbaren Freezer eingefroren. Drei Frischsamenparameter (Motilität, osmotische Resistenz und Haltbarkeit unter Laborbedingungen) und zwei Auftausamenparameter (Motilität und Kopfkappenintegrität) wurden erhoben und korreliert. Dabei enwies sichder osmotische Resistemtest als für die Beurteilung von Frischsperma hinsichtlich TG-Eignung gut geeignet, während die beiden anderen Frischsamenparameter keine bzw. nur schwache Korrelation zu den Auftauergebnissen aufwiesen. Aus den Auftausamenparametern wurde eine “Einfrierbarkeitszahl” als Maβ für die Eignung des Spermas zur Tiefgefrierkonservierung der einzelnen Eber bzw. Ejakulate erstellt. Dabei enwies sich der osmotische Resistenztest als für die Beurteilung von Frischsperma hinsichtlich TG-Eignung gut geeignet. Bei einem orientierenden Besamungsversuch wurden zufriedenstellende Abferkelraten erzielt.     

Use of a real-time polymerase chain reaction-based fluorogenic 5' nuclease assay to evaluate insect vectors of Corynebacterium pseudotuberculosis infections in horses

OBJECTIVE: To develop and use a sensitive molecular assay for detecting the phospholipase D (PLD) exotoxin gene of Corynebacterium pseudotuberculosis in an attempt to identify insect vectors that may be important in transmission of clinical disease in horses. SAMPLE POPULATION: 2,621 flies of various species. PROCEDURE: A real-time polymerase chain reaction (PCR)-based fluorogenic 5' nuclease (TaqMan) system (ie, TaqMan PCR assay) was developed for the detection of the PLD gene in insects. Flies were collected monthly (May to November 2002) from 5 farms in northern California where C. pseudotuberculosis infection in horses is endemic. Three of the 5 farms (which housed a total of 358 horses) had diseased horses during the study period. A total of 2,621 flies of various species were tested for the PLD gene of C. pseudotuberculosis. RESULTS: Evidence of bacterial DNA for the PLD gene was detected in skin biopsy specimens from clinically affected horses and from 3 fly species collected from farms where affected horses were housed. Farms with a high incidence of diseased horses had a high proportion of insects carrying the organism. High percentages of flies with positive results for the PLD gene were observed in October, when most clinically affected horses were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Our results are consistent with the hypothesis that C. pseudotuberculosis may be vectored to horses by flies. Three potential vectors were identified, including Haematobia irritans, Stomoxys calcitrans, and Musca domestica. The organism can be identified in up to 20% of house flies (Musca domestica) in the vicinity of diseased horses.     

Improved immunogenicity of novel baculovirus-derived Theileria parva p67 subunit antigens

East Coast fever (ECF) in cattle is caused by the tick-borne protozoan parasite Theileria parva. The major sporozoite surface antigen of T. parva (p67) is an important candidate for inclusion in a subunit vaccine. Recently, we reported the expression and production of different parts of p67 as fusions to either GFP or to the baculovirus GP64 envelope glycoprotein in insect cells, which resulted in stable proteins recognized by a monoclonal specific for native p67. The immunogenicity of these fusion proteins was examined in out-bred mice and cattle. In mice, the full length p67 molecule without its signal peptide and transmembrane region, but fused to GFP (GFP:p67deltaSS) was the best immunogen followed by the C-terminus of p67 fused to GP64 (GP64:p67C). These two immunogens also provoked a high level of sero-conversion in cattle when formulated in a water-in-oil or saponin-derived adjuvant with only 100 microg of protein and a single booster. The vaccine-elicited antibodies efficiently inhibited the infectivity of T. parva sporozoites in in vitro neutralization assays. This study demonstrated that these new baculovirus-derived p67 vaccines were highly immunogenic, and that in combination with a suitable adjuvant, they have a clear potential to induce protective immunity in cattle.     

Feline dental resorptive lesions in the 13th to 14th centuries

The Schild excavation (1971-1975) unearthed 1871 feline bones from at least 181 cats from the town market in medieval Schleswig-Gottorf. Seven of the 189 mandibles and one of the 126 skulls were investigated using a combination of macroscopic, radiographic, and histologic examinations as well as Knoop hardness measurements. The preliminary results of examinations of three mandibles and one skull are presented and reveal that feline dental resorptive lesions were present in cats that lived in a settlement period dating from the 13th and 14th centuries in former Schleswig, Germany.     

Toxic effects of mebendazole at high dose on the haematology of red-legged pademelons (Thylogale stigmatica)

OBJECTIVE: To investigate the effects of mebendazole at high dose on the haematology of macropods. Experimental. PROCEDURE: Five red-legged pademelons (Thylogale stigmatica) were dosed orally with mebendazole at 50 mg/kg/d for 5 to 6 days. Two control pademelons were dosed with water. Regular blood samples were taken for haematology over 20 days. RESULTS: All four treated pademelons sampled at 5 days developed severe leucopenia and neutropenia, moderate lymphopenia, thrombocytopenia, eosinopenia and monocytopenia, as well as bone marrow aplasia within 5 to 11 days after the first mebendazole dose. Four pademelons died unexpectedly or became ill and were euthanased 5 to 11 days after the first dose while the other animal recovered after 5 days of illness. Necropsy revealed systemic infection with opportunistic enteric bacteria, non-suppurative inflammation in tissues, haemorrhage and ulceration of the gastrointestinal tract. CONCLUSIONS: Red-legged pademelons rapidly develop bone-marrow aplasia and subsequent septicaemia after administration of high doses of mebendazole. Mebendazole at high doses should not be used for macropods.     

Molecular differentiation of Mycobacterium bovis isolates. Review of main techniques and applications

Until recently, none of the Mycobacterium bovis typing techniques permitted a satisfactory differentiation of isolates. During the last 10 years, the genome of pathogenic mycobacteria has been extensively studied, and phylogenetic analyses have shown that all (except Mycobacterium avium) belong to a single genetic species: the Mycobacterium tuberculosis complex. This increase in knowledge about the genome of these bacteria has lead to the discovery of molecular markers that allow us to differentiate isolates. Because of the phylogenetic proximity of the strains, even if most of these markers have been discovered in M. tuberculosis, they could be successfully adapted to the other bacteria of the M. tuberculosis complex, especially M. bovis. The most common markers in use today are the IS6110 insertion sequence, the direct repeat (DR) region, the poly(GC) rich (PGRS) sequences and the variable number tandem repeats (VNTR) sequences. The corresponding typing techniques are briefly described, and current knowledge of polymorphism and marker stability is detailed. If molecular markers are to offer wide perspectives for field studies, these two characteristics (polymorphism and stability) must be taken into account when choosing the marker(s) used in a study. In this context, examples of the application of molecular typing techniques for M. bovis are reviewed, on the one hand with epidemiological studies for which the major problem is the comparison between isolates and, on the other, with more general studies about the population genetics of M. bovis in a given country, and about its history and its phylogeny.     

Closure of large patent ductus arteriosus with a self-expanding duct occluder in two dogs

Of the different catheterisation methods described for closure of patent ductus arteriosus (PDA), coil embolisation is most commonly used in dogs. However, for a PDA larger than 4 to 5 mm in diameter, coil implantation is difficult. For these cases, the Amplatzer duct occluder (ADO) offers an alternative method. This report describes the successful implantation of an ADO in two dogs with large PDAs of approximately 6 mm diameter. The self-expandible device attached to an implantation wire was advanced through a long sheath antegrade to the femoral vein through the right heart and pulmonary artery to the duct and delivered into the PDA. Thereafter the device was released by unscrewing it from the delivery cable. The large PDA in both dogs was totally occluded by these means without any residual shunt. Thus, the ADO is a controlled release implant that also allows occlusion of a large PDA. Its high costs limit its general use in veterinary medicine at the present time.     

Efficacy, safety, and residue evaluation of levamisole gel formulation in sows

Anthelmintic efficacy, safety, and residue studies were conducted in sows and gilts with a levamisole gel containing 11.5% levamisole HCl. In 12 sows and 12 gilts, 8 mg of levamisole HCl equivalent/kg of body weight orally was 100% (resinate) and 91.1% (gel) effective against 55-day-old Ascaris suum and 100% (gel) and 96.1% (resinate) effective against Oesophagostomum dentatum. In 20 sows given levamisole gel (8 mg of levamisole HCl/kg) orally just before breeding, 4 to 6 weeks after breeding, 4 to 6 weeks before farrowing, and just before farrowing, there were no adverse effects. Transient salivation was noticed in five sows after treatment. In 4 groups of 4 sows each given levamisole gel orally to provide 8, 24, 40, or 80 mg of levamisole HCl/kg, adverse clinical signs were not observed in sows treated with 8 mg/kg. Transient salivation was noticed in one sow given 24 mg/kg, two sows given 40 mg/kg, and four sows given 80 mg/kg. Multiple emesis and chomping occurred in one sow given 80 mg/kg. Levamisole residues in edible tissues from sows given 8 mg of levamisole gel/kg orally were less than 0.1 mg/kg of muscle and fat in sows killed on posttreatment day (PTD) 3 and less than 0.1 mg/kg of kidney in sows killed on PTD 5. Liver residues averaged 0.78 mg/kg in sows killed on PTD 3 and were reduced to 0.31 mg/kg in sows killed on PTD 5. The 99% upper tolerance limit with 95% confidence on the withdrawal time to assure levamisole residues of less than 0.10 mg/kg in liver tissue was 11 days.