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Observation on the schizont stage of an unidentified Theileria sp. in experimentally infected sheep.

The schizont stage of an unidentified Theileria sp. infective for small ruminants was observed. Intact sheep were infected with adult Haemaphysalis qinghaiensis collected from fields where ovine theileriosis was prevalent. When the infested sheep developed theileriosis, tissue and organ smears were prepared. Theileria schizonts were demonstrated in liver, spleen, lung, kidney, lymph node and peripheral blood. Most of the schizonts were found outside the host cells, which was probably a smear artifact. This study adds our knowledge that lung and kidney can be parasitized by schizonts of Theileria sp.

Animals↗

Expression of the nucleocapsid protein of Dugbe virus and antigenic cross-reactions with other nairoviruses.

The small (S) RNA segment of Dugbe (DUG) virus (Nairovirus, Bunyaviridae) encodes a single protein, the nucleocapsid (N) protein, of M(r) 49.4 kDa. cDNA derived from the complete coding region for the N protein was cloned into Autographa californica nuclear polyhedrosis virus (AcNPV) under control of the polyhedrin promoter and used to infect Spodoptera frugiperda insect cells. Western blotting analysis using monoclonal antibodies demonstrated the production of DUG N protein in the infected cells. Monoclonal and polyclonal antibodies to the N protein of Crimean-Congo haemorrhagic fever (CCHF) virus were found to cross-react weakly with the baculovirus expressed DUG N protein by Western blotting. When used in an enzyme linked immunoassay (ELISA), the DUG N protein reacted with polyclonal mouse immune ascitic fluids raised against either CCHF or Hazara viruses (both members of the CCHF serogroup of nairoviruses). Cross-reactions between DUG virus (Nairobi sheep disease serogroup) and members of other nairovirus serogroups were not detected.

Antigens, Viral↗

Comparative nitric oxide production by LPS-stimulated monocyte-derived macrophages from Ovis canadensis and Ovis aries.

Bighorn sheep are more susceptible to respiratory infection by Mannheimia haemolytica than are domestic sheep. In response to bacterial challenge, macrophages produce a number of molecules that play key roles in the inflammatory response, including highly reactive nitrogen intermediates such as nitric oxide (NO). Supernatants from monocyte-derived macrophages cultured with M. haemolytica LPS were assayed for nitric oxide activity via measurement of the NO metabolite, nitrite. In response to LPS stimulation, bighorn sheep macrophages secreted significantly higher levels of NO compared to levels for non-stimulated macrophages. In contrast, levels of NO produced by domestic sheep macrophages in response to M. haemolytica LPS did not differ from levels detected in non-stimulated cell cultures. Nitrite levels detected in supernatants of LPS-stimulated bighorn macrophage cultures treated with an inducible nitric oxide synthase (INOS) inhibitor, N(G)-monomethyl-L-arginine, were similar to that observed in non-stimulated cultures indicating a role for the iNOS pathway.

Antigens, CD↗

Time profile of abamectin and doramectin excretion and degradation in sheep faeces.

We studied abamectin and doramectin excretion and their degradation in sheep faeces under field conditions on pasture after a single subcutaneous dose (0.2mg/kg body weight). In the excretion experiment, maximal abamectin concentration (1277 ng/g dry faeces) was detected on day 3, while doramectin concentration showed two peaks (2186 and 1780 ng/g dry faeces on days 2 and 5, respectively). Both avermectins were excreted at approximately the same rate (k=0.23 day(-1) for abamectin and 0.19 day(-1) for doramectin). In the field, a rapid loss of abamectin and doramectin from sheep faeces was seen during the first 32 days after which concentrations remained constant at approximately 77 ng/g and 300 ng/g, respectively. The half life values (DT(50)) for abamectin and doramectin dissipation from sheep faeces were 23 and 22 days, respectively, during the first 32 days. Dissipation of both avermectins was strongly correlated with moisture content of the faeces.

Animals↗

Host isolation and patterns of genetic variability in three populations of Teladorsagia from sheep.

We have used a mitochondrial marker to explore the population genetics of an economically important parasite of sheep, Teladorsagia. We examined diversity within and between parasites from three very different host populations, as well as within and between individual hosts. One of our study populations, the Soay sheep on Hirta, St Kilda, is unusually isolated with no sheep having been introduced to the island since 1932. Worm haplotypes from Hirta were compared with those from two other host populations. Remarkably, despite its historical isolation the Hirta population shows similar levels of within-population diversity to the other study populations. No divergence between the three Teladorsagia populations was found, consistent with gene flow between the populations. The high diversity within Teladorsagia populations provides compelling evidence that this variability is a general feature of parasitic nematode populations. Such diversity may be caused by high effective population size, coupled with an increased mutation rate for mtDNA, which has important implications for the spread of anthelmintic resistance in nematode populations.

Animals↗

The consequences of short-term grazing of bioactive forages on established adult and incoming larvae populations of Teladorsagia circumcincta in lambs.

The objective of this study was to investigate the consequences of short-term grazing on bioactive forages on (i) the viability and fecundity of established adult Teladorsagia circumcincta population and (ii) the establishment and development of incoming T. circumcincta infective larvae. Forty-eight, parasite naive, 3-month old, grazing lambs were artificially infected with 8000 infective larvae of T. circumcincta on day 1 of the experiment. On day 21 p.i., lambs were allocated to one of three bioactive forage grazing treatments; chicory (Cichorium intybus), sulla (Hedysarum coronarium), lotus (Lotus pedunculatus), and the control grass/clover (Lolium perenne/Trifolium repens) forage. On day 28 of the experiment a second dose of 8000 T. circumcincta infective larvae was administered to the lambs to investigate the effects of forages on the ability of infective larvae to establish within the host. All animals were slaughtered for worm recovery on day 35, while liveweight gain, feacal egg counts (FEC) and total worm egg output were monitored regularly throughout the experiment. Although FEC or total egg output were similar among the groups, adult worm burdens at slaughter were significantly affected (P<0.05) by forage treatment during the 2 week grazing period. Lambs grazing chicory had the lowest adult worm burdens and significantly lower numbers of male worms compared to those grazing on grass/clover (P<0.01), while the lambs grazing on sulla or lotus had similar adult populations to grass/clover fed animals. The results from the worm recoveries of the second dose (immature worm burdens) were affected by physiologically and/or immunologically mediated mechanisms, which reduced larval establishment in all treatments. Nevertheless, immature worm burdens at slaughter were similar between chicory, sulla and grass/clover group, while the immature worm recoveries from the lotus group were significantly higher (P<0.05) compared to those from lambs grazing grass/clover. Overall, the results of the present study support the view that chicory can be a promising candidate species in pasture management practices to control T. circumcincta burdens.

Animal Feed↗

Bluetongue virus detection by two real-time RT-qPCRs targeting two different genomic segments.

The detection of the bluetongue virus (BTV) by conventional methods is especially difficult and labour-intensive. Molecular diagnosis is also complex because of the high genetic diversity between and within the 24 serotypes of BTV. In the present study, two laboratories joined forces to develop and validate two new RT-qPCRs detecting and amplifying BTV segments 1 and 5. The 2 assays detect strains from all 24 serotypes. They both have a detection limit of 0.01 ECE50 and all 114 samples from BTV-free goats, sheep and cattle were negative. The two assays resulted in similar C(t) values when testing biological samples collected in sheep infected experimentally with a field strain of BTV from the Mediterranean basin. On average, the C(t) values obtained with the 2 methods applied to the 24 serotypes were not significantly different from each other, but some moderate to high differences were seen with a few strains. Therefore these two methods are complementary and could be used in parallel to confirm the diagnosis of a possible new introduction of BTV. An RT-qPCR amplifying a fragment of the beta-actin mRNA was also developed and validated as internal control for the bluetongue specific assays. The three assays described allow a reliable and rapid detection of BTV.

Animals↗

Molecular detection and characterization of Dichelobacter nodosus in ovine footrot in India.

Dichelobacter nodosus was detected in three clinical cases of ovine footrot in Kashmir, India. The detection was done by PCR in three clinical specimens directly, without isolating the organism, using species-specific 16S rDNA primers. Positive results were indicated by amplification of a 783 bp product. All the three samples were subjected to serogrouping by multiplex PCR using group (A-I) specific primers. All the three samples revealed the presence of serogroup B of D. nodosus by yielding a single band of 283 bps.

Animals↗

Identification of macrophage induced genes of Corynebacterium pseudotuberculosis by differential fluorescence induction.

Corynebacterium pseudotuberculosis is the etiological agent of the sheep disease caseous lymphadenitis. We have developed a promoter reporter system for this organism based on expression of the green fluorescent protein (gfp) gene from Aequorea victoria. A promoterless vector, pSM20, containing the gfp gene was constructed, and promoters were inserted upstream of the gfp gene. Upon transformation into C. pseudotuberculosis, fluorescence could be visualised by fluorescence microscopy, and relative promoter strength measured by flow cytometry. The usefulness of this system for measuring changes in gene expression was demonstrated by measuring fluorescence levels of heat shocked C. pseudotuberculosis carrying a dnaK promoter construct. Replication of C. pseudotuberculosis within J774 macrophages could be monitored by fluorescence microscopy. The establishment of the system allowed the use of differential fluorescence to identify genes that showed up-regulation following macrophage infection. Genes coding for a non-ribosomal peptide synthetase and the beta chain of a propionyl CoA carboxylase were identified as possessing promoters that demonstrated enhanced activity following macrophage infection by C. pseudotuberculosis.

Corynebacterium pseudotuberculosis↗

Crimean-Congo hemorrhagic fever virus genome L RNA segment and encoded protein.

Sequence analysis of the L RNA genome segment and predicted encoded L polymerase protein of Crimean-Congo hemorrhagic fever (CCHF) virus (genus Nairovirus, family Bunyaviridae) demonstrates that they are approximately twice the size of those found in viruses of other bunyavirus genera. The CCHF virus L segment and encoded protein (12,164 nucleotides and 3944 amino acids, respectively) are similar in size and sequence to those of the nairovirus Dugbe virus (12,255/62% and 4036/62% nucleotide and amino acid length/identity, respectively). The identification of an ovarian tumor (OTU)-like protease motif in the L protein amino termini of the nairoviruses Dugbe, CCHF, and Nairobi sheep disease (NSD) indicates these proteins are members of the recently described OTU-like protease family and suggests that these large proteins may be polyproteins that are autoproteolytically cleaved or involved in deubiquitination.

Amino Acid Motifs↗

National prevalence of larval echinococcosis in sheep in slaughtering plants Ovis aries as an indicator in control programmes in Uruguay.

Diagnostic surveys carried out on ovine larval echinococcosis in slaughtering plants at a national level in 1998 made it possible to meet several objectives, i.e. to estimate the existing levels of prevalence, to evaluate the effectiveness of the control actions applied by the Programme against Hydatidosis and to contribute to the improvement of the health information systems for epidemiological surveillance. The work included sheep (Ovis aries), both lambs (2-4 teeth) and adults (6-8 teeth). A random sample was performed among sheep from the plants that slaughter the majority of animals in the country. The sample included 22 animals per herd, and it involved removing their offal (liver, lungs and heart) and processing serial sections at the laboratory. The histopathological studies were performed on the lesions that could not be adequately identified at gross examination, and those that were so small that had to be submitted for confirmation of the diagnosis. Of the 2035 animals that were examined, 1019 were lambs (2-4 teeth) and 1016 were adults (6-8 teeth). The prevalence of larval echinococcosis was 7.7% in lambs and 18.0% in adults. The overall prevalence in the liver was 8.5 and 8.0% in the lungs. As to the characteristics of the morphologic structures of the larvae, in younger animals 29% of the lesions were calcified, and 71.0% were hyaline, while in older animals the proportions were 34.4% calcified versus 65.6% hyaline. Other conditions found in the liver parenchyma included: white stain (0.8%), caseous lymphadenitis (3.0%), abscess (0.4%), larvae of Taenia hydatigena (5.1%), Fasciola hepatica (3.9%). And 1.7% of the animals presented larvae of Taenia ovis in the cardiac muscle. Previous surveys carried out in 1994 using the same methodology showed a prevalence of 15.2% in 2-4 teethed animals and 33.9% in 6-8 teethed animals, thus confirming that rates have decreased by 49.6 and 47.0%, respectively. The data obtained reflect the reduction of areas contaminated with eggs of Echinococcus granulosus, which would cause a loss in immunity against this parasite after a certain time. Prevalence of larvae T. hydatigena and T. ovis could change if more sites were considered, a fact that was confirmed in studies carried out in our country. The species O. aries must be used as an indicator in the control programmes, especially in the category of 2-4 teethed lambs. The fact they are not as frequent in our market must be considered; in 1998 they accounted for only 3.0% of the total numbers of animals.

Abattoirs↗

The effect of xylazine epidural anaesthesia on blood gas and acid-base parameters in rams.

The blood gas and acid-base effects of epidural xylazine were studied in rams. Xylazine at a dose of 0.4 mg kg-1 led to a metabolic alkalosis with a significant increase of pH, plasma bicarbonate and base excess and respiratory depression. Epidural administration of xylazine provided prolonged and very effective analgesia during abdominal and hindlimb surgery in rams.

Acid-Base Equilibrium↗