Role of haemophilus pleuropneumoniae lipopolysaccharide endotoxin in the pathogenesis of porcine Haemophilus pleuropneumonia

Intact Haemophilus pleuropneumoniae cells (strain Shope 1, serotype 1), highly purified lipopolysaccharide (LPS) obtained from this strain of H pleuropneumoniae, as well as from Escherichia coli O111:B4, filter-sterilized H pleuropneumoniae cell-free culture supernatant fluid, and heat-inactivated supernatant fluid were given intranasally to CF1 mice and intratracheally to pigs. Pulmonary lesions induced by H pleuropneumoniae in mice were similar to those induced by H pleuropneumoniae in pigs. Histologically, lungs of mice and pigs killed 1 or 2 days after inoculation with 200 micrograms of highly purified H pleuropneumoniae LPS had lesions similar to one another and were similar to those in mice and pigs given intact H pleuropneumoniae, except that little or no necrosis or hemorrhage was observed. In mice killed 1 or 2 days after inoculation of 200 micrograms of E coli O111:B4 LPS, pulmonary lesions were similar to those in mice given H pleuropneumoniae LPS. Pulmonary lesions in mice given cell-free culture supernatant fluid obtained from a midlog-phase growth culture of H pleuropneumoniae cultivated in a chemically defined medium were severe and consisted of neutrophil infiltration and extensive necrosis. In mice, the heat-inactivated supernatant fluid produced mild lesions that consisted of foci of neutrophil aggregation and no necrosis. Extensive necrosis observed in lesions caused by cell-free culture supernatant fluid could be attributed to the action of a heat-labile component, perhaps by the extracellular heat-labile hemolysin produced by H pleuropneumoniae cultivated in chemically defined medium. A LPS endotoxin and a heat-labile factor may be involved in the pulmonary lesion development in the acute phase of porcine Haemophilus pleuropneumonia.     

An experimental model for subclinical edema disease (Escherichia coli enterotoxemia) manifest as vascular necrosis in pigs.

An experimental model for subclinical edema disease was developed in weanling pigs. In multiple experiments, 3-week-old pigs were weaned, then inoculated intragastrically with 10(10) colony-forming units of an SLT-IIv-positive strain of Escherichia coli originally isolated from a pig with edema disease (principals). Control pigs were inoculated with a nonpathogenic E coli strain. Of 39 principals, 8 developed clinical edema disease within 14 days after inoculation. However, 20 of 21 principals that did not develop clinical signs of edema disease, but were submitted for necropsy examination at 14 days after inoculation, had characteristic vascular lesions of edema disease. Vascular lesions, found principally in ileum and brain, consisted of segmental necrosis of myocytes in the tunica media of small arteries and arterioles. None of the pigs inoculated with a nonpathogenic strain of E coli developed edema disease or vascular lesions. None of the principals necropsied at 2 days after inoculation had vascular lesions. Development of vascular lesions by 14 days after inoculation was used as the end point for detecting subclinical edema disease in the model.     

Detection with nonradioactively labeled probes of the toxin genes of different E. coli pathotypes from swine]

We tested hemolytic E. coli from 86 pigs with edema disease or colidiarrhea. They were tested serologically and with nonradioactive digoxigenin-dUTP labelled probes for the presence of enterotoxin or Shiga-like-toxin genes. By slide-agglutination we detected 38 cases with E. coli O149:K88, 28 with E. coli O139:82B and 20 with E. coli O141. E. coli of serogroup O149:K88 isolated from diarrheic pigs, reacted with the probes for LT and STb genes. Edema disease E. coli O139:82B reacted with the SLTII probe. E. coli O141, isolated from colidiarrhea or edema disease showed a diversity of toxin gene patterns. All the E. coli O141 from diarrheic pigs reacted with the probes for LT and STap in addition to SLTII. No strains isolated from pigs with edema disease possessed any of these enterotoxin genes. Gene probe technique confirmed the serological method as useful tool for diagnosing E. coli O149:K88 and O139:82B as ETEC or VTEC, respectively. On the other hand only the demonstration of toxin genes with probes could explain the pathological findings in the pigs shedding E. coli of serogroup O141.     

Characteristics of verotoxigenic Escherichia coli from pigs.

Porcine verotoxigenic Escherichia coli were characterized with respect to frequency of occurrence, serogroup, and association with disease, weaning, and selected properties of the bacterium. Of 668 strains of E. coli from southern Ontario pigs with enteric disease, 32 (4.8%) produced verotoxin at 10(3)-10(7) cytotoxic doses per mL of culture supernatant. Of 22 isolates which belonged to O serogroups 138, 139 and 141, 15 produced verotoxin. Among other enterotoxigenic types of E. coli, two of 57 isolates of O157:K"V17" and two of 96 isolates of O149:K91 were verotoxigenic. The remaining 13 verotoxigenic E. coli belonged to O groups 2, 107, 120, 121 and 130. An additional 21 verotoxigenic E. coli belonging to O groups 138, 139 and 141 and three to O157:K"V17" were identified in a collection of 47 E. coli recovered from weaned pigs with enteric disease. Verotoxigenic E. coli were associated with postweaning diarrhea, bloody stools, sudden death and edema disease. They were isolated at similar frequencies (14%) from healthy weaned pigs, and from weaned pigs with enteric disease. Isolation rates from neonates were low and significantly different from rates in weaned pigs. Neutralizing antibody to verotoxin was not detected in the sera of 45 pigs, which included pigs from herds with a history of edema disease. Verotoxin was not associated with production of colicin, hemolysin, or enterotoxins or with any of 23 biochemical properties of the organisms. The serological data indicate that porcine verotoxigenic E. coli are not a common source of verotoxigenic E. coli for humans. Porcine verotoxin may play a role in postweaning diarrhea and absence of detectable neutralizing antibody in serum may be an important aspect of pathogenesis.     

Treatment of Edema Disease of Swine

Antihistamine (Allermin) was used sucessfully in the treatment of naturally occurring edema disease of swine under the field and laboratory conditions. A high recovery rate (88.8%) was obtained in the group of pigs with mild clinical signs. The intravenous route of Allermin administration had definite beneficial value. The release of histamine induced by circulating immune-complexes may be responsible for the clinical signs characteristic of edema disease.

Three pigs that recovered clinically following the Allermin treatment were examined histologically. Fibrinoid persisted in the vascular walls at least 30 days after the initiation of the treatment. The fibrinoid vasculitis, generalized eosinophilia at the tissue level, and favorable response of pigs to antihistamine treatment suggest that edema disease may be caused by allergy.

     

Reproduction of edema disease of swine with purified Shiga-like toxin-II variant.

Twenty-one weaned Yorkshire-Landrace pigs were injected intravenously with graded doses of purified Shiga-like toxin-II variant (edema disease toxin). In a preliminary study, three pigs (Nos. 1, 2, 3) were injected with 48, 24, and 12 ng, respectively, of SLT-IIv/kg of body weight. Subsequently, three groups (Nos. 4, 5, 6) of six pigs each were injected with 6, 3, and 1.5 ng, respectively, of SLT-IIv/kg of body weight. Severe clinical signs and histologic lesions characteristic of edema disease developed in pigs Nos. 1, 2, and 3, and all six pigs in group No. 4. Eight of these pigs were euthanatized in extremis (mean time to death was 34 hours) and one died of the disease (52 hours). Moderate signs and lesions of edema disease were observed in all pigs in group No. 5, and three pigs were euthanatized (mean time to euthanasia was 42 hours). Mild signs and lesions were observed in three pigs in group No. 6. The most common gross pathologic changes were edema of the eyelids, submucosa of the stomach, and mesentery of the spiral colon and hemorrhage of the colon and cerebellum. Microscopic lesions were associated with vascular injury and included vessel necrosis, perivascular edema and hemorrhage, and superficial colonic and cecal erosions. The vascular lesions were observed in the cerebellar folia, submucosa and mucosa of the stomach, cecum, colon, and sporadically in the retina. None of the clinical signs associated with endotoxin were observed.     

Vascular ultrastructure and DNA fragmentation in swine infected with Shiga toxin-producing Escherichia coli

Shiga toxins (Stx) produced by Escherichia coli cause systemic vascular damage that manifests as edema disease in swine and hemolytic uremic syndrome in humans. In vitro, Stx inhibit protein synthesis and, depending on circumstances, induce necrosis, apoptosis, or both. The mechanism of in vivo Stx-mediated vascular damage is not known. The ability of Stx to cause apoptosis of vasculature in vivo was studied in pigs with edema disease that was produced by oral inoculation with Stx-producing E. coli. Arterioles of ileum and brain were evaluated by terminal dUTP nick-end labeling (TUNEL) assay for DNA fragmentation in myocytes (10 infected pigs, 5 control pigs) and by transmission electron microscopy for ultrastructural changes characteristic of apoptosis (17 infected pigs, 8 control pigs). In comparison with controls, increased numbers of TUNEL-positive arterioles were detected in 6/10 (60%) subclinically affected pigs 14-15 days after inoculation. Ultrastructurally, lesions in myocytes consisted of lysis (necrosis), with cytoplasmic debris and nuclear fragments contained between intact basement membranes. Endothelial cell changes ranged from acute swelling to necrosis and detachment from basement membrane. Subclinically affected pigs (n = 14) tended to have changes predominantly in myocytes, whereas pigs with clinical illness (n = 3) more commonly had changes in endothelial cells. The arteriolar lesions and clinical signs of edema disease are attributed to the effects of Stx on vasculature. Therefore, our findings suggest that the Stx-induced arteriolar lesions seen in this study were primarily necrotic, not apoptotic. We suspect that necrosis was the principal cause of the DNA fragmentation detected.     

仔猪水肿病病原菌的分离鉴定

从江苏淮安某猪场具有典型水肿病临床症状和剖检变化的发病猪组织分离的菌株中挑选5株大肠杆菌,分别命名为JS0001、JS0002、JS0003、JS0004和JS0005。通过培养特性、菌体形态、染色特性、生化鉴定及血清学试验等一系列方法对其进行系统鉴定,并对这5株细菌进行抗药性、溶血性、Vero细胞毒性和小白鼠致病性等试验。试验结果表明,JS0001、JS0002、JS0004和JS0005表现β型溶血,腹腔接种小白鼠,JS0001、JS0002、JS0003、JS0004对其具有较高致病性;5株细菌药敏特性基本一致：对痢特灵（FR）、头孢噻肟（CTX）、阿米卡星（AN）敏感,对头孢曲松（CRO）敏感性最高,而对阿莫西林(AMX)、多西环素(DO)等药物具有耐药性。JS0002、JS0004能够分泌Vero细胞毒素,确定为产类志贺毒素大肠杆菌（SLTEC）。血清学试验结果表明,JS0002、JS0004的血清型为O₁₃₉,JS0001、JS0003为O₁₄₁。     

致仔猪水肿病大肠杆菌的分离鉴定及对小鼠的毒力研究

从发病仔猪脏器中分离到一株细菌,进行了初步鉴定。结果表明,该菌为致猪水肿病大肠杆菌,在血平板上呈β溶血,经PCR鉴定该菌株具有产毒素基因,对小鼠毒力强,能致其发病死亡,血清型为O141,是临床多发血清型菌株。将细菌培养物和裂解毒素分别经皮下和尾静脉两种途径接种于体重为16~18 g昆明系小鼠,小鼠腹腔注射细菌培养物后12~48 h死亡,不表现神经症状,剖检水肿病病变特征不典型;小鼠于尾静脉注射裂解毒素后4~48 h,出现典型的眼睑水肿和后躯瘫痪等神经症状,直至死亡。该菌株致小鼠死亡的最小毒素剂量为0.2 mL/只。研究结果表明,小鼠可作为诊断仔猪水肿病模型,也进一步证明试验菌株为致猪水肿病菌株,可作为制备疫苗菌株。     

Expression of cyclooxygenase-2 in swine naturally infected with Actinobacillus pleuropneumoniae

Cyclooxygenase-2 (COX-2) was detected and localized in 15 pigs with naturally occurring pleuropneumonia using a 437-base pair digoxigenin-labeled cDNA probe in an in situ hybridization protocol. Histopathologic changes in the acute stage were characterized by coagulative necrosis of lung parenchyma, hemorrhage, vascular thrombosis, edema, fibrin deposition, and infiltration of lung parenchyma by neutrophils and alveolar macrophages in nine pigs. In chronic lesions, a thick layer of granulation tissue surrounded foci of pulmonary necrosis in six pigs. All 15 pigs infected with Actinobacillus pleuropneumoniae, confirmed by bacterial isolation, had distinct positive hybridization signals for COX-2 in bronchial, bronchiolar epithelial cells, alveolar macrophages, neutrophils, and type I pneumocytes. COX-2 expression was detected primarily in neutrophils from pigs with acute lesions and primarily in alveolar macrophages from pigs with chronic lesions. The results suggest that a prostanoid product of COX-2 is an important component of the inflammatory response to acute and chronic A. pleuropneumoniae infection.     

Effects of Escherichia coli Shiga-like toxins (verotoxins) in pigs.

Escherichia coli K12 strains TB1(pCG5), with the genes for Shiga-like toxin IIv from an edema disease isolate of E. coli and TB1(pCG5-1), with the toxin genes inactivated by transposon mutagenesis, were used to test the hypothesis that Shiga-like toxin IIv was the same as edema disease principle. Ammonium sulfate precipitated culture supernatants from the pair of E. coli K12 strains and from a wild edema disease isolate of E. coli (E145) were tested for their ability to induce signs and lesions of edema disease in intravenously inoculated weaned pigs. Similar preparations from E. coli which produce Shiga-like toxins I and II were also tested. Preparations from E. coli TB1 (pCG5) and E145 contained high levels of Shiga-like toxin IIv and induced signs and lesions similar to those seen in edema disease, whereas preparations from E. coli TB1 (pCG5-1) failed to induce signs or lesions of edema disease. All Shiga-like toxin preparations produced delayed neurological signs, fibrinoid necrosis of arterioles and hemorrhages in the cerebellum of pigs. High doses of Shiga-like toxin IIv were associated with superficial necrosis of the colonic epithelium and vasculitis. Shiga-like toxins I and II resulted in kidney lesions but no enteric pathology. Shiga-like toxin II preparations had the lowest median lethal dose for pigs, Shiga-like toxin IIv was intermediate and Shiga-like toxin I was the least toxic.     

Prevention of edema disease in pigs by vaccination with verotoxin 2e toxoid.

Pigs in 2 herds with persistent problems with post weaning edema disease caused by infection with verotoxin-2e (VT2e)-producing Escherichia coli O139 were treated with a VT2e-toxoid vaccine. Treatment was performed as a randomized blind field trial with parallel treatment and non-vaccinated control groups. In 1 herd, a group of pigs was injected with adjuvant alone. Pigs were vaccinated at 1 and 3 wk of age and weaned at 4 wk of age. The effect of vaccination was measured by average daily weight gain (ADG), mortality due to edema disease within the 1st 4 wk after weaning, and weight at 3-6 mo of age. Pathological and microbiological examinations were performed on all pigs that died during the 1st 4 wk post weaning. Only pigs from which VT2e+, F18+ E. coli O139 was isolated were categorized as "death due to edema disease." The serological response to vaccination was evaluated by an indirect ELISA. Vaccination had a statistically significant effect on the level of antibodies specific for VT2e in both herds. Vaccination resulted in a statistically significant increase in ADG in the nursery period but not in the grower-finishing period. Vaccination had a statistically significant effect on mortality due to edema disease with an odds ratio of 0.039, indicating that there was almost total elimination of mortality due to the disease in the vaccine groups.     

Dermal responses to Ostertagia ostertagi in Ostertagia ostertagi- and Cooperia punctata-inoculated calves

Calves harboring patent Ostertagia ostertagi or Cooperia punctata were given intradermal injections of O ostertagi 3rd-stage larval antigen. The initial injections were followed 30 days later by a 2nd series of injections. Skin thickness was measured at injection sites for 72 hours after injection. Selected injection sites including saline solution control sites were biopsied at 30 minutes, at 3, 24, 48, and 72 hours, and at 30 days after injection. After the 1st series of injections, there was a clear distinction in dermal reactions between O ostertagi-inoculated calves and C punctata-inoculated calves; after 24 hours, reactions were not seen in the C punctata-inoculated calves. Marked dermal reactions occurred in the O ostertagi-inoculated calves. The reactions at 30 minutes and 3 hours were characterized by slight-to-extensive infiltration of neutrophils and dermal edema. The 24-hour cellular reaction was principally due to neutrophil and eosinophil infiltration with edema and necrosis. Reactions at 48 to 72 hours were due to eosinophils and perivascular accumulations of macrophages and lymphocytes. Necrosis, neutrophils, and edema were present in foci where fragments of nematodes were located. On reinjection, a clear distinction in dermal reactions between calves was not seen based on the type of nematode infection. Thirty days after dermal inoculation, large nodules developed at the site of the initial antigen injection. The nodules were characterized by marked intradermal proliferation of lymphocytes in a follicular pattern with occasional macrophages and rare multinucleated giant cells.     

Virulence profiles of Shiga toxin 2e-producing Escherichia coli isolated from healthy pig at slaughter

Recently, virulence patterns of Stx2e-producing Escherichia coli from pigs with edema disease and from humans were compared and strains from diseased pigs were reported to be unlikely human pathogens [Sonntag, A.K., Bielaszewska, M., Mellmann, A., Dierksen, N., Schierack, P., Wieler, L.H., Schmidt, M.A., Karch, H., 2005. Shiga toxin 2e-producing Escherichia coli isolates from humans and pigs differ in their virulence profiles and interactions with intestinal epithelial cells. Appl. Environ. Microbiol. 71, 8855-8863]. In the present study, 31 Shiga toxin-producing E. coli (STEC) strains harboring stx2e, which were previously isolated out of fecal samples from healthy pigs at slaughter [Kaufmann, M., Zweifel, C., Blanco, M., Blanco, J.E., Blanco, J., Beutin, L., Stephan, R., 2006. Escherichia coli O157 and non-O157 Shiga toxin-producing Escherichia coli in fecal samples of finished pigs at slaughter in Switzerland. J. Food Prot. 69, 260-266], were characterized by phenotypic and genotypic traits. Nine of the thirty-one sorbitol-positive non-O157 STEC (stx2e) isolated from healthy pigs belonged to serotypes found in STEC isolated from humans, including two serotypes (O9:H-, O26:H-) reported in association with hemolytic-uremic syndrome. Otherwise, the serotypes were different from those isolated from cases of edema disease in pigs. The eae (intimin) gene, which is strongly correlated with severe human disease, was not detected. Moreover, all strains were lacking the genes for enterohemolysin (ehxA), porcine A/E associated protein (paa), STEC autoagglutinating adhesin (saa) and the serin protease EspI (espI). Nine strains tested positive for astA (EAST1), one O141:H17 strain for fedA (F18 fimbrial adhesin) and one O159:H- strain for terF (tellurite resistance). Similar to the Stx2e-producing E. coli isolated from humans, which are mainly lacking further virulence factors, genes of an iron uptake system on the high-pathogenicity island (irp2, fyuA) were detected in three ONT:H10 and ONT:H19 strains from healthy pigs. Consequently, although the isolated strains are unlikely to be associated with severe human diseases, healthy pigs cannot be excluded as a potential source of human infection with Stx2e-producing STEC.     

Experimental Edema Disease of Swine (E. Coli ENTEROTOXEMIA) I. Detection and Preparation of an Active Principle

Freeze-thaw lysates prepared from strains of Escherichia coli belonging to serogroups O138, O139, and O141 contained a principle (edema disease principle) which induced edema disease in swine. All freeze-thaw lysates contained endotoxic activity that tended to obscure the edema disease syndrome and methods were developed to reduce such activity. Freeze-thaw lysates prepared from E. coli O139 induced the most characteristic edema disease syndrome. Partially purified edema disease principle prepared from O139 freeze-thaw lysates by sequential precipitation with ammonium sulphate and streptomycin sulphate had increased specific activity with markedly reduced endotoxic activity. This material was insoluble at acidic pH but readily soluble at alkaline pH. The effective molecular weight of edema disease principle, based on retention and filtration properties of diaflo membranes, appeared to be greater than 50,000 and less than 100,000. The biological activity of edema disease principle was thermolabile. Sodium deoxycholate treatment of edema disease principle further reduced endotoxic activity. A thermolabile, ammonium sulphate precipitable material was prepared from E. coli O139 that induced a predictable syndrome which resembled edema disease clinically and pathologically following intravenous inoculation in pigs.     

Flunixin meglumine attenuation of endotoxin-induced damage to the cardiopulmonary vascular endothelium of the pony

Endotoxic shock was induced in 5 ponies by intraperitoneal injections of 20, 40, 60, 80, and 80 micrograms of Escherichia coli endotoxin (LPS)/kg of body weight at 0, 6, 12, 18, and 24 hours, respectively. At 24 hours, the ponies also were given 20 micrograms of LPS/kg via catheter in the left ventricle of the heart. A 2nd group of 4 ponies was given 1.1 mg of flunixin meglumine (FM)/kg, IV, at 6, 12, 18, and 24 hours just before the corresponding LPS injection. Two hours after the 24-hour LPS injection, the ponies in both groups were anesthetized, the lungs were perfused with fixative, and portions of the pulmonary arteries and veins and right and left ventricles were prepared for scanning and transmission electron microscopy. In ponies that were given only LPS, some areas of pulmonary vascular endothelium appeared normal when compared with untreated controls, but other areas had disoriented endothelial cells or had varying amounts of sloughing, which ranged from focal areas of a few cells to large areas of denuded endothelium. Ponies treated with FM before LPS had less severe and less extensive endothelial cell damage. In both groups, leukocytes were attached to areas of the vessel wall; endothelial cell damage was greater in these regions. Administration of FM before LPS administration attenuated the LPS-induced endothelial cell damage.     

Escherichia coli strains from edema disease: O serogroups, and genes for Shiga toxin, enterotoxins, and F18 fimbriae

The objectives of the research were to determine the presence of the gene sequences for Shiga Toxin 2e (Stx2e), enterotoxins (ST-I, ST-II and LT-I), and F18 fimbriae in 144 Escherichia coli strains isolated from pigs with edema disease; to assess the ability of stx2e(+) strains to produce Stx2e; and to determine the O serogroups of the E. coli strains. Presence of the genes was determined by polymerase chain reaction (PCR), production of Stx2e was assessed by cytotoxicity for Vero and Hela cells, O serogroups were identified by agglutination with specific antisera. Of the 144 strains tested, 99 were stx2e(+) by PCR, but only 45 of these were Stx2e(+) in the cell culture assays. Among the 99 stx2e(+) strains, PCR detected the genes for F18ab, ST-I, ST-II, LT-I in 76, 40, 31 and 16 strains, respectively. Forty-one of the 99 sxt2e(+) strains belonged to O group 139; the rest did not belong to the classical edema disease O serogroups. It is likely that the enterotoxins, whose genes were detected at high frequency, are responsible for diarrhea seem in pigs with edema disease in Brazil.     

Phenotyping of E. coli serotypes associated to oedema disease

Background

Oedema disease is a severe disease, mainly affecting recently weaned pigs. It is caused by E. coli strains that express fimbriae F18 and produce verotoxin 2e, mainly belonging to serotype O138, O139 or O141. The aim of this study was to compare E. coli isolates within these serotypes with respect to diversity.

Methods

Faecal E. coli strains belonging to serotypes O138, O139 and O141 isolated during the period 1994–1998 from Swedish pigs aged less than 12 weeks were compared using a biochemical fingerprinting system. Aiming to compare the results obtained over time, also strains isolated during 1964–67 and 1975–80 were included in the study. The study comprised 129, 263 and 95 isolates of E. coli serotype O138, O139 and O141, respectively.

Results

Biochemical phenotypes (BPTs) were defined. At each sampling occasion each herd could only contribute with one isolate per BPT. Consequently, all but one of identical BPTs identified at a specific sampling occasion was omitted. The final number of isolates from 1994–98 that was compared included 64, 182 and 41 isolates of serotypes O138, O139 and O141, respectively. Within each serotype, the dominating BPT included over 65% of the compared isolates, demonstrating a large dominance of one BPT per serotype. These dominating BPTs were also demonstrated in the material from the 1960ies and the 1970ies. Still, the presence of other common BPTs (especially within serotype O138 and O139) demonstrated a certain variation within serotype. In a herd severely affected by oedema disease, E. coli serotype O139 was easily demonstrated in diseased pigs but only rarely in apparently healthy weaners

Conclusion

The results obtained demonstrate the presence of dominating BPTs within the oedema disease inducing serotypes. A stability of these BPTs over time was observed, presumably at least partly due to a never-ending access to naïve pigs. Still, the presence of other common BPTs indicates a variation over time, which visualises the importance of monitoring for this. Such studies should focus on pigs affected by oedema disease, because oedema disease inducing strains of E. coli were only rarely demonstrated in healthy pigs in a herd affected by oedema disease.     

An induced synovitis disease model in ponies

The effects of intra-articular injection of small amounts of E. coli lipopolysaccharide (LPS) into the intercarpal joint of 5 ponies were studied. The LPS induced predictable changes all of which were analogous to acute bacterial infection, except that the development of signs occurred sooner after the LPS injection, and subsided within 36 hours. Fever was monophasic and peaked at 5-7 hours. The ponies exhibited depression, reduced or absent appetite, increased pulse and respiration rates, and lameness. The lameness became evident between 1 and 2 hours after injection, at which time warmth, articular effusion, and resentment to palpation of joint flexion were evident. Hematological changes included neutrophilic leucocytosis, and changes in copper, iron and zinc serum concentrations. The synovial fluid total protein, leucocyte, and alkaline phosphatase levels increased within 2 hours. The mucin precipitation, total protein and leucocyte counts in synovial fluid remained elevated long after clinical and hematological changes had subsided. The model is useful for the study of some aspects of infectious joint disease.     

Effects of Cold Compression, Bandaging, and Microcurrent Electrical Therapy after Cranial Cruciate Ligament Repair in Dogs

Objective— To compare 4 therapeutic techniques to reduce soft tissue swelling after cranial cruciate ligament repair in the dog.
Study Design— Prospective study.
Animals— Twenty-four dogs with cranial cruciate ligament rupture (CCLR).
Methods— Dogs with naturally occurring CCLR, were surgically repaired by an extracapsular technique and randomly divided into 4 treatment groups (cold compression [CC], modified Robert Jones bandage [B], cold compression and bandage [CCB], and microcurrent electrical therapy and bandage [METB]) each with 6 dogs. Data were collected at 2 time points, the morning after surgery before the 1st treatment and 72 hours later after the last treatment. Limb girth was measured at 3 anatomic locations to assess soft tissue swelling and all affected limbs were evaluated for presence (or absence) of pitting edema and bruising. Analysis of covariance was used to determine effect of treatment on the percent change in circumference. Duncan's multiple-range test was used to determine differences in treatment groups circumferential percent change over 72 hours. Statistical significance was set at P <.05.
Results— Use of a Robert Jones bandage had the least effect on reducing postoperative soft tissue swelling with CC, METB, and CCB being equally effective in reducing swelling by 72 hours after surgery.
Conclusion— Use of cold compresses alone or with a bandage, or using microcurrent electrical therapy in combination with a bandage decreases soft tissue swelling over 72 hours more than a bandaging alone after extracapsular repair of CCLR.
Clinical Relevance— CC, METB, and CCB should be considered as viable options to limit soft tissue swelling after extracapsular repair of CCLR in dogs.