The O-antigen of Salmonella enterica serotype Enteritidis PT4: a significant factor in gastrointestinal colonisation of young but not newly hatched chicks

The lipopolysaccharide of Salmonella and other Gram negative pathogenic species has been implicated as a major virulence determinant and in this study we report the role of LPS of S. Enteritidis in the colonisation and persistent gastrointestinal infection of young poultry. The gene encoding the unique O-antigen ligase, waaL, was mutated by insertional inactivation in a well characterised S. Enteritidis strain, S1400/94. The waaL mutant, designated PCP, produced rough colonies on agar medium, did not agglutinate O9 antiserum, did not produce an LPS ladder on silver stained gels and was serum sensitive. PCP and a nalidixic acid marked derivative of S1400/94 (S1400/94 Nalr) were used to orally challenge young chicks, separately and together in competitive index experiments. At post-mortem examination of 1-day-old chicks challenged S1400/94 Nalr and PCP separately there were no significant differences in the numbers of S1400/94 Nalr and PCP bacteria in tissues sampled on days 1, 2, and 5. By day 42 after challenge S1400/94 Nalr bacteria were recovered in significantly higher numbers than PCP from the caecal contents (P < 0.001). In competitive index studies in the 1-day-old chick PCP colonised, invaded and persisted in lower numbers than S1400/94 Nalr. In 4-week-old chicks challenged separately, PCP bacteria were recovered from all tissues examined in significantly lower numbers than S1400/94 Nalr. In competitive index experiments in 4-week-old chicks, PCP was not detected at any site and at any time point. Therefore, the O-antigen of S. Enteritidis plays an important role in poultry infections although this role is less important in the newly hatched chick.     

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.     

Pancreatic exocrine secretion and plasma concentration of some gastrointestinal hormones in response to abomasal infusion of starch hydrolyzate and/or casein

Eight Angus steers (290 +/- 8 kg), surgically prepared with pancreatic pouch-duodenal reentrant cannulas and abomasal infusion catheters were used in a replicated 4 x 4 Latin square experiment to investigate the effects of abomasal infusion of starch hydrolyzate (SH) and/or casein on pancreatic exocrine secretion and plasma concentration of hormones. Steers were fed a basal diet of alfalfa (1.2 x NEm) in 12 equal portions daily. Abomasal infusion treatments (6-L total volume infused per day) were water (control), SH [2.7 g/(kg BW x d)], casein [0.6 g/(kg BW x d)], and SH + casein. Periods were 3 d for adaptation and 8 d of full infusion. Pancreatic juice and jugular blood samples were collected over 30-min intervals for 6 h on d 11. Weight and pH of pancreatic samples were measured, and a 10% subsample was composited and frozen until analysis of total protein and pancreatic enzyme activities. The remaining sample was returned to the duodenum. Plasma was harvested and frozen until analyzed. Pancreatic juice (67 mL/h) and protein (1.8 g/h) secretion rates were not affected by nutrient infusion. There were SH x casein interactions for all pancreatic enzyme secretions (U/h; alpha-amylase, P < 0.03; trypsin, P < 0.08; and chymotrypsin, P < 0.03) and plasma insulin concentration (P < 0.10). Secretion of pancreatic enzymes was increased by SH (trypsin) and casein (alpha-amylase, trypsin, and chymotrypsin) but not when SH + casein were infused together. Glucose (P < 0.10) and cholecystokinin octapeptide concentrations (CCK-8; P < 0.05) were increased by SH, but glucagon was decreased (P < 0.10). Casein decreased (P < 0.10) plasma CCK-8 concentrations. These data indicate that positive effects of postruminal casein on enzyme secretion were inhibited by SH, emphasizing the complexity of the regulatory mechanisms involved in dietary adaptation of pancreatic exocrine secretion. Changes in hormone concentration may not relate directly to changes in enzyme secretion.     

Comparison of bupivacaine femoral and sciatic nerve block versus bupivacaine and morphine epidural for stifle surgery in dogs

ObjectiveTo evaluate the efficacy of combined femoral and sciatic nerve blocks as an alternative to epidural anesthesia and analgesia in dogs undergoing stifle surgery under general anesthesia.Study designProspective, blinded, randomized, clinical comparison.AnimalsTwenty dogs weighing 37 ± 11 (mean ± SD) kg, aged 3 (1–8) [median (minimum–maximum)] years undergoing elective unilateral tibial-plateau leveling osteotomy.MethodsDogs were assigned randomly to receive either epidural anesthesia (bupivacaine 0.5%, 0.5 mg kg^?1 + morphine 0.1%, 0.1 mg kg^?1, in 0.2 mL kg^?1; EPID) or femoral and sciatic nerve blocks (Bupivacaine 0.5%, 0.1 mL kg^?1, was administered at each site; F + S) guided by electrolocation. All patients received a standard general anesthesia technique. Pain and sedation were scored (on scales of 0–10 and 0–3, respectively) pre-operatively, at extubation, and at 1, 4 and then every 4 hours thereafter up to 24 hours. Postoperatively, hydromorphone was administered to any patient with a pain score of >5 or whenever the blinded caregiver determined that more hydromorphone was necessary. Intraoperative heart rate (HR), mean arterial pressure (MAP), end tidal isoflurane (FE′ISO), body temperature, post-operative pain scores, time to first hydromorphone dose after surgery, time to first feeding, time to first drinking, time to first urination, time to first ambulation (walk on a lead) and cumulative dose of hydromorphone were recorded.ResultsIntra-operatively, FE′ISO and MAP were significantly lower in the EPID group (p = 0.05 and p = 0.04, respectively). Postoperatively, the cumulative hydromorphone consumption (p = 0.04) and the incidence of urinary retention (p = 0.03) were higher in the EPID group.Conclusion and clinical relevance F + S is a practical alternative to EPID that produces less urine retention and reduces opioid consumption in the 24 hours after surgery. EPID might be associated with a lower isoflurane requirement and lower systemic blood pressure.     

Subcutaneous Botryomycosis Due to Bibersteinia trehalosi in a Texas Longhorn Steer

A 3-year-old Texas Longhorn steer had a long history of progressive swelling of the soft tissues of the jaw and neck. At necropsy, multifocal to coalescing dermal and subcutaneous pyogranulomas were surrounded by fibrous tissue. Microscopically, the pyogranulomas contained aggregates of gram-negative coccobacilli surrounded by Splendore-Hoeppli material and were separated by bands of fibrovascular tissue (botryomycosis). Phylogenetic analysis of multilocus sequence-typing data revealed that the bacteria recovered in pure culture from swabs of submandibular tissue were most closely related to Bibersteinia [Pasteurella] trehalosi. The bacterial colonies were immunohistochemically reactive with a rabbit polyclonal anti-Pasteurella class C acid phosphatase antibody. Botryomycosis is a pyogranulomatous inflammation caused by a variety of nonbranching, nonfilamentous bacteria that elicit the formation of Splendore-Hoeppli material. This case of botryomycosis is unique for its association with Bibersteinia trehalosi.     

Multifocal cutaneous histiocytic sarcoma in a young dog and review of histiocytic cell immunophenotyping

A 9‐month‐old male Great Dane had progressive generalized nodular dermatopathy for several months. There were > 100 raised, alopecic, firm, painful nodules throughout the skin. Aspirates from several lesions yielded moderate numbers of irregularly round or polygonal to spindle‐shaped cells with mild to moderate anisocytosis and few inflammatory cells, and the cytologic interpretation was proliferation of mesenchymal or histiocytic cells. On histopathologic examination, nodules were composed of densely packed sheets of round to spindle‐shaped cells with mild anisokaryosis and low mitotic activity. Multifocal histiocytic sarcoma with a spindle‐cell pattern was diagnosed based on morphologic features and intense expression of CD18. Additional immunophenotypic analysis on frozen sections of tissue confirmed the diagnosis of histiocytic sarcoma; expression of CD18, CD45, CD1a, CD11b, and CD11c, limited expression of Thy‐1 (CD90) and CD80, and lack of expression of CD4, CD11d, and CD86 indicated that the cells were likely interstitial dendritic cells; a review of reactive and neoplastic dendritic cells is provided. Based on staging, internal organs were not affected. Sequential treatment with lomustine and doxorubicin failed to prevent progression of the cutaneous lesions, and the dog died 3 months after initial diagnosis. At necropsy, a focus of neoplastic cells was present in one lymph node, but except for skin other organs were not involved. The clinical presentation of histiocytic sarcoma may be unusual, and neoplastic cells may lack overt features of malignancy on cytologic and histopathologic examination. In some instances, immunophenotyping is required to differentiate histiocytic sarcoma from other histiocytic disorders.     

Index selection in terminal sires improves early lamb growth

The use of terminal sires (TS) for crossbreeding is integral to the UK sheep industry where approximately 71% of market lambs are sired by TS rams. Early growth of these crossbred lambs affects profitability. The objectives of this study were i) to evaluate the effectiveness of index selection among TS on BW and ADG of their crossbred offspring; and ii) to compare the efficacy of that selection within TS breeds. The most widely used TS breeds in the United Kingdom are Charollais, Suffolk, and Texel. These participated in sire referencing schemes in which they were evaluated on a lean growth index designed to increase carcass lean weight at a given age. From 1999 to 2002, approximately 15 high and 15 low lean growth index rams per breed (93 in total, differing in index on average by 4.6 SD) were selected from within their sire referencing schemes and mated to Welsh and Scottish Mule ewes. Their crossbred offspring were reared commercially on 3 experimental farms in England, Scotland, and Wales. A total of 6,515 lambs were born between 2000 and 2003. Lambs were weighed at birth (BWT), 5 wk (5WT), and 10 wk (10WT), and their ADG from birth to 10 wk was calculated. Lambs sired by high index rams were on average, across breeds, heavier at all ages (P < 0.01) with 0.07 ± 0.03, 0.3 ± 0.1, and 0.4 ± 0.1 kg greater BWT, 5WT, and 10WT, respectively. Their ADG was 5.1 ± 1.9 g/d greater than low-index-sired lambs (P < 0.01). Suffolk-sired lambs were on average heavier at all ages, with greater ADG, whereas Charollais-sired lambs were lightest with smallest ADG. Overall, there was no significant interaction between sire index and sire breed (P > 0.10). Within Suffolk-sired lambs, there was little difference between high and low index sires for the traits studied (P > 0.3). High and low index Charollais-sired lambs differed in BWT (0.09 ± 0.04 kg) and 5WT (0.3 ± 0.1 kg), and Texel-sired lambs differed in 5WT (0.5 ± 0.1 kg), 10WT (0.9 ± 0.2 kg), and ADG (10.2 ± 3.3 g/d; P < 0.01). Lambs from Scottish Mule dams were heavier, with greater ADG, than lambs born to Welsh Mules (P < 0.01). Lambs reared in Scotland were heavier at all ages (P < 0.01). The results suggest that using index selection in TS can improve the growth of their commercial offspring reared on grass.     

Morphology and Neurochemical Expression of Neurons Immunoreactive for the Calcitonin Gene-Related Peptide (CGRP-IR) in the Lamb Ileum

Veterinary Research Communications -     

Cardiorespiratory Effects of Combined Midazolam and Butorphanol in Isoflurane-anesthetized Cats

The cardiorespiratory effects of midazolam (0.1 mg/kg) and butorphanol (0.4 mg/kg) were evaluated in seven cats. Cats were induced in a chamber, removed and intubated, and maintained with isoflurane in 100% oxygen. Cardiorespiratory data were recorded before and 3, 10,20,30,40,50, and 60 minutes after intravenous injection of midazolam-butorphanol. Heart rate, mean arterial pressure, and respiration rate were significantly (p < .05) decreased below baseline at various times throughout the study. Tidal volume and minute ventilation were unchanged. End-tidal carbon dioxide was significantly (p < .05) increased above baseline for 30 minutes. Midazolam-butorphanol administered intravenously induced significant alterations in several cardiorespiratory parameters in isoflurane-anesthetized cats.