The detection and characterisation of Dichelobacter nodosus from cases of ovine footrot in England and Wales

Footrot, caused by the strictly anaerobic bacterium Dichelobacter nodosus, is the most common cause of lameness in sheep in Great Britain but problems exist in association with its diagnosis and control. The fastidious nature of D. nodosus means that complex media and several weeks are required for characterisation. An alternative method to simplify and enhance the detection of D. nodosus in clinical samples is therefore highly desirable. In terms of control, anecdotal evidence from the farming community suggests that the commercially available vaccine, based on Australian isolates of D. nodosus, is not widely employed in this country due to its perceived inefficacy. Seven hundred and six isolates, collected from outbreaks in England and Wales, were therefore used to investigate these issues. A 16S rRNA PCR was adapted to detect D. nodosus in clinical material within 1 day of sampling; a 15% increase in detection compared with culture and less than 1% false negatives were achieved. This represents a major advance in the rapid diagnosis of footrot and will be of great value to practitioners and diagnostic laboratories. Bacterial virulence was tested using protease thermostability and zymogram assays, whilst serogrouping was performed by slide agglutination. All isolates demonstrated virulence patterns previously recorded in Australia and all nine serogroups of D. nodosus (A-I) were represented. Serogroup H was predominant. There was, therefore, no evidence for the presence of novel strains of D. nodosus compared with Australia suggesting the need for further investigation into farmers' views on the use of the commercial vaccine in Great Britain.     

Infarctive purpura hemorrhagica in five horses

Five horses were examined because of signs of muscle stiffness, colic, or both. All 5 had been exposed to Streptococcus equi within 3 weeks prior to examination or had high serum titers of antibodies against the M protein of S equi. Horses had signs of unrelenting colic-like pain and focal areas of muscle swelling. Four horses were euthanatized. The fifth responded to treatment with penicillin and dexamethasone; after 3 weeks of treatment with dexamethasone, prednisolone was administered for an additional 10 weeks. Common hematologic and serum biochemical abnormalities included neutrophilia with a left shift and toxic changes, hyperproteinemia, hypoalbuminemia, and high serum creatine kinase and aspartate transferase activities. Necropsy revealed extensive infarction of the skeletal musculature, skin, gastrointestinal tract, pancreas, and lungs. Histologic lesions included leukocytoclastic vasculitis in numerous tissues and acute coagulative necrosis resembling infarction. These horses appeared to have a severe form of purpura hemorrhagica resembling Henoch-Sch?nlein purpura in humans and characterized by infarction of skeletal muscles. Early recognition of focal muscle swelling, abdominal discomfort, neutrophilia, hypoalbuminemia, and high serum creatine kinase activity combined with antimicrobial and corticosteroid treatment may enhance the likelihood of a successful outcome.     

The management and welfare of some common ovine obstetrical problems in the United Kingdom

This review approaches sheep welfare during the periparturient period from a veterinary practitioner viewpoint. At the most fundamental husbandry level, the general hygiene standards adopted on many sheep farms in the United Kingdom (UK) during the lambing period are unacceptable and require urgent improvement. Concerns also arise from obstetrical conditions attended by farmers which cause pain that can be identified using clinical criteria such as altered demeanour and behaviour of the ewe, and the presence of inflammation of the reproductive tract. Reduced production is evidenced by poor milk production and hungry lambs, the necessity to rear such lambs artificially and, in neglected cases, death of the ewe and her lambs. The correction of dystocia, periparturient vaginal, uterine and rectal prolapses is greatly facilitated after epidural injection of lignocaine, or a combination of xylazine and lignocaine administered by a veterinary surgeon. However, the perceived high professional fees charged for these services, and for Caesarean operations, result in few sheep receiving veterinary care, with such neglect directly contributing to poor welfare standards and an increased ewe mortality rate during the periparturient period. Whilst ending any welfare concern, the destruction of ewes rather than requesting veterinary services purely for economic reasons is a disturbing trend in sheep production in the UK and raises concerns regarding positive interaction between veterinary practitioner and farmer. In this article, veterinary treatments which provide effective analgesia are described and costed for obstetrical conditions in sheep. Recommended treatment protocols are described for the common obstetrical problems in sheep. In addition to appropriate treatment of obstetrical problems and alleviation of pain, if veterinary involvement could reduce ewe periparturient mortality from estimated figures of 4-2% this would save an estimated 400,000 ewes annually in the UK. Sheep welfare could be considerably improved by practising veterinarians providing practical instruction to small groups of clients with funding provided by government agencies.     

Acute post-parturient haemoglobinuria in dairy cows and phosphorus status

During the conduct of an experiment designed to examine the nutritional management of dairy cows in late pregnancy, four cows out of 72 suffered from acute haemoglobinuria two to four weeks after calving. Thirty-six thin and 36 fat cows were individually fed one of three diets based on a total mixed ration with different energy or protein concentrations during the last 3 to 4 weeks before expected calving date. After calving, cows grazed pasture and were offered 6 kg dry matter of pelleted concentrates daily. The P concentrations of the feeds offered suggested that the cows' diets were marginally deficient in P relative to requirements. Plasma P concentrations were significantly (P < 0.05) lower in fat cows than in thin cows during the first 6 weeks of lactation (0.87 versus 1.12 mmol/L), but precalving diet had no effect (P > 0.05). Concentrations of plasma inorganic P of the four fat cows that developed acute haemoglobinuria were less than 0.3 mmol/L. However, plasma P concentrations in another 12 cows, none of which displayed overt symptoms, declined to similar levels. It appeared that inadequate dietary P may have predisposed cows to acute haemoglobinuria, but the precipitating cause was not readily obvious.     

Concentration of pro-vitamin A carotenoids in common beef cattle feedstuffs

Quantification of the pro-vitamin A carotenoids in feedstuffs commonly fed to livestock has been ignored for many years. A greater dietary concentration of vitamin A has the potential to limit adipogenesis in cattle, thereby reducing carcass quality and value. A survey of 18 feedstuffs commonly fed to beef cattle was conducted for determination of vitamin A equivalents based on analysis of carotenoids. The pro-vitamin A carotenoids of interest were β-carotene, α-carotene, and β-cryptoxanthin. Collaborators in 5 states collected the feedstuffs and then shipped them to The Ohio State University for compilation and analysis. Carotenoids were extracted from the feedstuffs and then quantified using HPLC with photodiode array analysis. Fresh fescue pasture contained approximately 10 times more vitamin A equivalents than hay and 5 times more than corn silage (39,865, 2,750, and 6,900 IU of vitamin A/kg of DM for fresh pasture, hay, and corn silage, respectively). Beta-cryptoxanthin and α-carotene could not be detected in any forage samples. Hay and corn silage vitamin A equivalents decreased over extended periods of time from harvest to sample collection. Corn was the only feedstuff to have appreciable concentrations of all 3 pro-vitamin A carotenoids quantified. Corn processing had a minimal impact on the vitamin A equivalents. High-moisture corn contained 54% more vitamin A equivalents than whole shelled corn (378 and 174 IU of vitamin A/kg of DM, respectively). Pro-vitamin A carotenoids were more concentrated in corn coproducts than in whole shelled corn. The drying of distillers grains with solubles may significantly degrade β-carotene (800 and 480 IU/kg of DM for wet and dry distillers grains, respectively). Soybean-based feedstuffs contain a small concentration of pro-vitamin A carotenoids, at 55 and 45 IU of vitamin A/kg of DM for soybean meal and soybean hulls, respectively. Overall, there was considerable variation in the pro-vitamin A content of feedstuffs based on location and storage conditions. An extensive analysis of feedstuffs would need to be conducted for an accurate estimation of the vitamin A content of feedlot cattle diets.     

Exposure to Sarcocystis spp. in horses from Spain determined by Western blot analysis using Sarcocystis neurona merozoites as heterologous antigen

Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs.     

Tumor necrosis factor-α and acute-phase proteins in early pregnant ewes after challenge with peptidoglycan-polysaccharide

Bacterial infection shortly after mating interferes with establishment of pregnancy. Injection of peptidoglycan-polysaccharide (PG-PS), a component of gram-positive bacteria, into sheep on day 5 after mating reduces pregnancy rate. Experiments were designed to evaluate the acute-phase response (APR) in ewes to injection of PG-PS on day 5 after mating (day 0). Catheters were inserted into the jugular and posterior vena cava on day 4. On day 5, ewes were challenged with saline or 30 μg/kg body weight (BW) PG-PS (Exp 1) or 60 μg/kg BW PG-PS (Exp 2). Blood samples were collected every 15 min for 6 h (Exp 1) and every 15 min for 2 h, hourly for 12 h, and at 24, 36, and 48 h (Exp 2). Body temperature and clinical signs of infection were monitored in Exp 2. Plasma was assayed for concentrations of a pro-inflammatory cytokine, tumor necrosis factor-α (TNF-α); 2 APR proteins, serum amyloid A (SAA) and haptoglobin (Hp); and progesterone (P₄). Ewes injected with 60 μg/kg BW PG-PS exhibited fever, vaginal discharge, loss of appetite, and lethargy. After challenge with either 30 μg/kg or 60 μg/kg BW PG-PS, TNF-α increased in the posterior vena cava. Concentrations of SAA and Hp in the jugular increased after challenge with 60 μg/kg BW PG-PS. Only half (5/10) of the ewes treated with 60 μg/kg BW PG-PS had ultrasonically visible embryos, and none of them had functional corpora lutea (CL) (<1 ng/mL of P₄) on day 21. On the other hand, 8/9 (88.9%) control ewes had visible embryos and all had functional CL on day 21. Using logistic regression, pregnancy on day 21 was predicted to depend on concentrations of TNF-α and Hp on day 5 and concentration of P₄ on day 14. In summary, injection of PG-PS on day 5 after mating resulted in fever; increased concentrations of TNF-α, Hp, and SAA on the day of and the day after the PG-PS challenge; and decreased concentrations of P₄ on days 14 and 21. These factors were related to failure to establish pregnancy.     

Inflammation-associated responses in piglets induced with post-weaning colibacillosis are influenced by dietary protein level

Forty piglets (average body weight = 5.32 kg) were used to investigate the effect of dietary crude protein (CP) content on immunological responses following a challenge with enterotoxigenic Escherichia coli (ETEC) K88. Pigs, housed 4 per pen, were randomly allotted to 2 diets: 1) a high, 225 g/kg CP diet (HCP) or 2) a low, 176 g/kg CP diet (LCP) supplemented with crystalline amino acids. Pigs were orally challenged with 6 mL of an ETEC K88 suspension containing 10¹⁰ cfu/mL on d 8 after weaning. Blood samples were collected from 10 pigs (1 pig/pen) on d 7 (at weaning), −24 h, 8 h, 72 h and 7 d after the challenge for determination of plasma urea N (PUN) and serum concentrations of tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1β) and haptoglobin (Hp). Tumor necrosis factor alpha, IL-1β and Hp were measured as indicators of inflammatory responses. The concentrations of serum TNF-α at 8 h, 72 h and 7 d after challenge were similar to the level observed at 24 h before challenge but higher (P < 0.05) than the weaning level. Pigs fed the LCP diet had lower (P = 0.032) concentrations of IL-1β (72 vs. 116 pg/mL) at 8 h post-challenge compared with those fed the HCP diet. Likewise, pigs fed the LCP diet tended to have lower (P = 0.088) concentration of Hp (9 vs. 25 mg/dL) compared with those fed the HCP diet at 8 h post-challenge. Compared with the weaning concentration, PUN concentration at 72 h after ETEC challenge was higher (P < 0.05) in pigs fed the HCP diet. The results indicate that the LCP diet supplemented with crystalline amino acids reduced inflammatory responses, as indicated by serum IL-1β, in piglets infected with ETEC K88.