Colonisation of the respiratory tract of lambs by strains of Mycoplasma ovipneumoniae

The age and time of year when colonisation of the nasal cavity of lambs by Mycoplasma ovipneumoniae occurs; the persistence of the organism, and its prevalence in the lungs at slaughter were examined in 2 flocks of sheep in New Zealand. No colonisation had occurred at the time of weaning at 6–7 weeks, but M. ovipneumoniae was recovered from most lambs on at least one occasion before they were slaughtered when about 8 months old. In most cases, colonisation of the nasal cavity by M. ovipneumoniae was a transient phenomenon. At slaughter M. ovipneumoniae was recovered from the lungs of 89% of the lambs of one flock and 80% of the other flock.

Bacterial restriction endonuclease DNA analysis (BRENDA) of 34 nasal isolates from one flock showed that it was possible to identify 7 “groups” each with markedly different BRENDA patterns. Lambs initially colonised by one strain, often lost that strain, and if recolonisation occurred it was with a different strain.

M. ovipneumoniae was recovered at slaughter from the lungs of most lambs, both normal and pneumonic. The isolates from one flock were examined by BRENDA, and approximately 90% of them gave similar or identical patterns. The predominant strain isolated from the lungs had been recovered from the nasal cavity of many of the lambs about 3 weeks earlier. This suggests that the nasal and lung isolates do not represent independent populations. However, nasal strains may differ in their ability to colonise the lungs.     

牛支原体NM001株单克隆抗体的制备与鉴定

为获得抗牛支原体NM001株单克隆抗体,并评价其特性,以牛支原体分离株NM001作为抗原免疫6周龄BALB/c小鼠,利用杂交瘤技术和间接ELISA方法筛选到2株能稳定分泌抗牛支原体的单克隆抗体细胞,命名为2C5和7G3。其细胞上清的间接ELISA抗体效价分别为6.4×10³和1.2×10⁴。经亚型测定,单抗2C5和7G3均属于IgG1类,轻链均是λ型。制备腹水并对单抗进行纯化和特性鉴定,两株单抗的间接ELISA抗体效价分别为1.02×10⁵和4.09×10⁵,且两株单抗与无乳支原体、山羊支原体山羊肺炎亚种、丝状支原体山羊亚种、绵羊肺炎支原体、牛巴氏杆菌均无交叉反应。Western Blot结果显示,2株单抗均能特异性识别牛支原体全菌蛋白中的相应蛋白。试验表明,单抗2C5和7G3能够与牛支原体发生特异性反应,从而为牛支原体血清学检测提供一定的物质基础。     

Seroepidemiologic study of natural transmission of Mycoplasma hyopneumoniae in a swine herd

A cohort of 57 pigs in a farrow-to-finish swine herd with mild clinical mycoplasmal disease was followed to determine patterns of seroconversion to Mycoplasma hyopneumoniae (MH), detected with an enzyme-linked immunosorbent assay (ELISA). Survival analysis was used to evaluate the relationship between time to seroconversion and possible risk factors for MH infection (or enzootic pneumonia).

Pigs were housed in outdoor pens at approximately 9 weeks of age, when passively acquired MH antibodies had decayed. From 9 to 11 weeks of age and during a 5 week period, pigs were exposed by direct (nose-to-nose) or indirect contact to older seropositive gilts. Blood samples were collected from each pig at 3 week intervals until market age, when they were either slaughtered or selected for breeding. Antibody concentration was measured as the ratio of optical densities of the serum sample to the positive control (S/P). Based on the sample distribution of S/P ratios from pigs in an MH-free herd, pigs were considered positive when S/P ratios were greater than 0.34. At the beginning of the study, all pigs were seronegative to MH. Seroconversion was first detected after 21 days, and was most frequent about 11 weeks after exposure to older seropositive gilts. By the end of the study, 11 pigs (19%) had seroconverted, with S/P ratios ranging from 0.40 to 1.11. The presence of gross lung lesions showed a moderate to good agreement with ELISA results (K = 0.62). Histologic lesions were evident in virtually all slaughtered pigs, ranging from mild, non MH-specific lesions to severe lesions typical of MH infection. No secondary respiratory pathogens were isolated. Clinical signs were mild and there was no significant difference (P > 0.4) in weight gain between seropositive and seronegative pigs, or between pigs with and without lung lesions. A Cox regression model was fitted to the seroconversion data, and opportunity of contact (direct or indirect) was the only significant variable. After adjustment for breed and antibody S/P ratio prior to exposure, pigs in direct contact with seropositive gilts were seven times more likely to seroconvert than those in only indirect contact.     

Kinetics of substrate oxidation and hydrogen peroxide production by Mycoplasma mycoides subsp. mycoides Large Colony (LC) type and Mycoplasma mycoides subsp. capri

Mycoplasma mycoides subsp. mycoides Large Colony (LC) type is a pathogen of goats causing contagious agalactia and respiratory disease, found on all continents where small ruminants are kept. It shares close genetic characteristics with M. mycoides subsp. capri. Substrate oxidation by 22 strains of M. mycoides subsp. mycoides LC from nine countries was compared with that of eight strains of M. mycoides subsp. capri from five countries. There was considerable similarity in the substrates used, but substrate saturation coefficients (K_s) varied for different substrates. Substrate utilization patterns and K_s values did not (1) significantly differentiate the LC strains from each other, (2) show any correlation with geographical origin, or (3) distinguish the LC strains from the capri strains. These results support previous studies justifying the reclassification of these subspecies as a single species.     

Detection of Mycoplasma mycoides sub-species mycoides small colony by a specific capture/enrichment monoclonal antibody-based sandwich ELISA

The present study describes the development of a specific Mycoplasma mycoides subsp. mycoides Small Colony (MmmSC) monoclonal antibody (MAb), 6E3, and its application in a sandwich ELISA (sELISA) format. Mab 6E3 reacted only to the 12 MmmSC within the 32 M. mycoides cluster strains and 12 representative strains of other bovine, ovine and caprine associated mycoplasmas examined. A capture/enrichment format of the sELISA that combined MAb 6E3 with a previously developed MAb 3H12 that cross reacted with Mmm Large Colony [Rodriguez, F., Ball, H.J., Finlay, D., Campbell, D., Mackie, D.P., 1996. Detection of Mycoplasma mycoides sub-species mycoides by monoclonal antibody-based sandwich ELISA. Veterinary Microbiology 51, 69–76], retained MmmSC specificity and improved the sensitivity from the 1.2 × 10⁷ cfu/ml for a standard 2 h capture stage sELISA down to as low as 2 cfu/ml for a 72 h capture. A low level of false positives (1%) was observed when this assay was applied to 200 bovine respiratory and milk samples submitted for diagnostic investigation. This simple and specific sELISA provides a suitable assay for screening large numbers of samples for CBPP.     

副猪嗜血杆菌外膜蛋白P5的B细胞抗原表位鉴定

副猪嗜血杆菌外膜蛋白P5能诱导机体产生免疫保护反应,同时可以用于特异性的血清学诊断,本试验选取P5蛋白进行抗原表位鉴定。首先通过PCR扩增P5F1（1～204aa）,P5F2（170～296aa）及P5F3（280～371aa）3个片段,PCR产物分别定向克隆到表达载体pET32a（＋）中表达纯化。根据ELISA和Western blotting结果确定P5F3片段（280—371aa）是OmpP5的免疫优势决定区。为了进-步对该免疫优势决定区进行抗原表住鉴定,设计了-套11个部分重叠的短肽,这些短肽覆盖全部280～371aa片段。每-个短肽合成1对寡核苷酸链,退火后插入表达载体pGEX-6p-1,与GST进行融合表达。用HPS阳性血清进行ELISA和Western blotting扫描,鉴定出其表位位于”。TGNTCDAVKGRKALIT351。通过序列分析证实该抗原表位在不同的HPS菌株中高度保守。本试验确定了位于HPSOmpP5上的-个抗原表位,为建立-种方便、快捷、适用于现地大规模样品检测的鉴别诊断方法奠定了基础,同时也为HPS新型亚单位疫苗的研制,以及研究病原茵感染和机体免疫过程中P5蛋白与宿主体内相应分子之间的相互作用提供了有用信息。     

Identification of an immunodominant 40 kDa merozoite antigen common to the Australian T and Dixie vaccine strains of Babesia bovis and the development of diagnostic tests specific for these strains

Antigenic differences among Australian vaccine and field strains of Babesia bovis were investigated in an attempt to identify strain specific antigens. Immunoblots revealed substantial differences between the current vaccine strains, designated T and Dixie, and previous vaccine strains and field isolates collected on properties where vaccination with the T or Dixie strains had failed to provide complete protection against tick-borne challenge. A major difference was an immunodominant 40 kDa antigen (T40) present in only the T and Dixie strains. The molecular weight and immunodominant nature of this antigen suggest that it may be the equivalent of the major merozoite surface antigen (MSA-1) described by others in North American strains of B. bovis. MSA-1 was shown to be conserved in north American isolates but not in an isolate from Israel or in the Australian S and L isolates. The work presented here suggests that merozoite surface antigen diversity exists among geographically different isolates of B. bovis within Australia.

Monospecific antiserum to T40 was used to develop an indirect fluorescent antibody (IFA) test specific for T and Dixie strain parasites, and a blocking enzyme-linked immunosorbent assay (ELISA) specific for antibody to the T and Dixie strains. In cases of babesiosis in recently vaccinated cattle, the IFA test will be a useful tool for determining whether clinical symptoms are due to a severe vaccine reaction or to a concurrent tick-borne infection. In a preliminary assessment of potential of the ELISA for the serological identification of vaccinated cattle using a total of 160 sera, the test clearly differentiated between animals vaccinated with the T or Dixie strains and non-vaccinated animals, and was not affected by presence of antibodies to other B. bovis strains.     

Immune responses to the capsular polysaccharide of Mycoplasma dispar in calves and mice

Humoral and cell-mediated immune responses to the capsular polysaccharide (CPS) of M. dispar and polygalacturonic acid (pGaIU—a structurally similar polysaccharide) were investigated in calves experimentally infected with Mycoplasma dispar and in mice immunized with CPS or pGaIU. Sera, tracheobronchial lavage and nasal fluids, collected before and after infection in calves, were checked for the presence of anti-CPS and anti-pGaIU antibodies. The sera from mice injected with CPS or pGaIU were checked for different classes of anti-CPS and anti-pGaIU antibodies. Peripheral blood lymphocytes from calves and splenic lymphocytes from mice were monitored for specific proliferative responses to CPS and pGaIU. At about 2 weeks post-infection, anti-CPS IgM response in serum, anti-CPS and anti-pGaIU IgM and IgA response in lavage fluid and lymphocyte proliferative response was seen in the calves. Mice immunized with CPS and pGaIU gave exclusively IgM responses. No secondary response was seen in mice immunized with CPS in contrast to mice immunized with pGaIU. Antibodies cross-reactive with pGaIU were present in the sera of CPS-immunized mice but antibodies cross-reactive with CPS were not found in pGaIU-immunized mice. No significant blastogenic response was shown by mouse splenocytes to CPS or pGaIU.     

Attenuation of a Brucella abortus mutant lacking a major 25 kDa outer membrane protein in cattle

OBJECTIVE: To determine the virulence of a Brucella abortus mutant, BA25, lacking a major 25 kd outer membrane protein (Omp25) in cattle. ANIMALS: 20 mixed-breed heifers in late gestation. PROCEDURE: 10 heifers were inoculated with 1 x 10(7) colony-forming units of the Omp25 mutant via the conjunctival sac, and an equal number were infected with the virulent parental strain B. abortus 2308. The delivery status of the dams was recorded, and colonization was assessed following necropsy. The ability of BA25 to replicate inside bovine phagocytes and chorionic trophoblasts was also evaluated in vitro because of the propensity of virulent brucellae to replicate inside these cells in vivo. RESULTS: The parental strain induced abortions in 5 of 10 inoculated cattle, whereas only 1 of 10 dams exposed to BA25 aborted. Brucella abortus strain 2308 colonized all of the cow-calf pairs and induced Brucella-specific antibodies in 100% of the dams. In contrast, BA25 was isolated by bacteriologic cultural technique from 30% of the calves and 50% of the inoculated dams (n = 10). Of the 10 heifers inoculated with BA25, 4 did not develop Brucella-specific antibodies nor were they colonized by the mutant strain. In bovine macrophages and chorionic trophoblasts, BA25 replicated in significantly lower numbers than the virulent parental strain (n = 3). CONCLUSIONS AND CLINICAL RELEVANCE: The 25 kd outer membrane protein may be an important virulence factor for B. abortus in cattle. The attenuation of the Omp25 mutant in cattle may involve the inability of BA25 to replicate efficiently in bovine phagocytes and chorionic trophoblasts.     

Serotype distribution and production of muramidase-released protein, extracellular factor and suilysin by field strains of Streptococcus suis isolated in the United States

Streptococcus suis is an important swine pathogen and a zoonotic agent. Differences in virulence have been noted among the 33 described serotypes, serotype 2 being considered the most virulent. In this study, we aimed at assessing the serotype distribution and the production of virulence-associated markers by strains recovered from diseased pigs in the United States (U.S.). Results showed that among the 100 strains evaluated, serotype 3 (20% of the isolates) and serotype 2 (17%) were the most prevalent. We then investigated the presence in these isolates of the genes sly, epf and mrp, encoding the virulence-associated markers suilysin (SLY), extracellular factor (EF) and muramidase-released (MRP) protein, respectively. The effective production of the markers by the strains was also verified. Results showed that the presence of the gene did not always correlate with actual expression of the respective protein. In the case of MRP, this was due, in most cases, to frameshift mutations at the 5′ end of the gene resulting in premature stop codons. The most prevalent phenotypes among U.S. strains were MRP⁺EF⁻SLY⁻ (40%) and MRP⁻EF⁻SLY⁺ (35%). Serotype distribution greatly differed from that reported in several European countries, as did the production of virulence markers, particularly for serotype 2. On the other hand, our results for the U.S. S. suis isolates are similar to those reported for Canadian strains, suggesting a common status in North America.     

8株鳖源变形杆菌外膜蛋白的比较

从 1 2批送检病鳖体内分离出 8株细菌 ,经形态学检查、生理生化特性测定、致病性测定和血清学鉴定 ,确定 7株为普通变形杆菌 ,1株为奇异变形杆菌。进一步采用十二烷基硫酸钠 (SDS)破菌法提取 8株鳖源变形杆菌的外膜蛋白 (OMP)进行SDS PAGE电泳 ,比较分析细菌OMP型。结果显示奇异变形杆菌的OMP型由 3条相对分子质量范围为 3 0× 1 0 4～4 3× 1 0 4的主要蛋白带组成 ;7株普通变形杆菌的主要外膜蛋白相对分子质量范围为 3 0× 1 0 4～6 7× 1 0 4,分属 2个OMP型 ,其中 4个分离株属OMP1型 ,由 4条主要蛋白带组成 ;其余 3个分离株属OMP2型 ,由 7条主要蛋白带组成。表明不同种变形杆菌的OMP型差异较大 ,同种不同株变形杆菌的OMP型相似 ,但蛋白带的迁移率及颜色深浅在菌株间仍有差异。此外 ,发现相对分子质量约为 4 3× 1 0 4的一条外膜蛋白带为所有菌株所共有 ,可能是变形杆菌属特异性抗原     

A microeconomic evaluation of the impact of Mycoplasma meleagridis infection in turkey production

A microeconomic evaluation approach was used to determine the economic impact of Mycoplasma meleagridis (MM) infection in turkeys. Profit maximization, using the production, cost and profit functions, and prices of inputs and outputs was utilized to determine the economically optimal time birds should be raised as well as the corresponding optimal level of feed input and liveweight meat output. Optimal was defined as that level which maximized profits.

Using regression analysis, the predicted weights of MM-infected (MM(+)) males and females were found to be significantly greater, for a given age or level of feed input, than noninfected (MM(−)) cohorts of identical age and parent line. This unusual weight advantage demonstrated by the infected birds may be the result of higher embryo mortality of this group thus reducing the survivability of some of the potentially poor weight gainers in the MM(+) group. By evaluating the profit functions, this weight advantage also translated into a profit advantage of $0.06 per bird associated with raising MM(+) turkeys.

Although the weight gain results were obtained from birds raised under environmentally controlled conditions and thus not subjected to typical overcrowding and climatic stress faced under field conditions, they agree with recent results obtained in a field study (Carpenter et al., 1982a). This apparent profit advantage should be considered when measuring the benefits derived from averting decreased hatchability, leg problems, and other anomalies associated with MM infection. This microeconomic approach should aid producers, veterinarians and administrators in their disease control/eradication decision making.     

Serum and mucosal antibody responses against Mycoplasma hyopneumoniae following intraperitoneal vaccination and challenge of pigs with M hyopneumoniae

Pigs were immunised intraperitoneally when six weeks old and again at about 10 weeks old with killed Mycoplasma hyopneumoniae antigen prepared in an oil adjuvant. The pigs were challenged with live M hyopneumoniae (Beaufort strain) at between 11 and 15 weeks old. Antigen specific antibody levels for both IgG and IgA classes in serum and respiratory tract secretion were monitored over time. In serum anti-M hyopneumoniae antibody was detected shortly after the second intraperitoneal vaccination and was largely IgG. In respiratory tract secretion the response was observed after challenge, and was primarily IgA. Anti-M hyopneumoniae antibody-containing cells and their immunoglobulin class specificity were monitored in lung and tracheal lamina propria. In lung the majority of anti-M hyopneumoniae-containing cells were IgG, whereas in the tracheal lamina propria the majority were IgA. These results are discussed in terms of the use of intraperitoneal vaccination for the control of M hyopneumoniae infection.     

禽多杀性巴氏杆菌血清型与外膜蛋白型相关性研究

为了解禽多杀性巴氏杆菌(Pasteurella multocida,Pm)分离菌株的荚膜血清型、菌体血清型与外膜蛋白型之间的相关性,首先对自行分离的10个菌株采用间接血凝试验和琼脂扩散试验进行鉴定(均为A:1);然后采用超声波破碎、高速离心和十二烷基肌氨酸钠提取外膜蛋白,通过SDS-PAGE电泳的方法对上述10个菌株与C48-1(A:1)、X73(A:1)、P1059(A:3)、CU(A:3,4)等一起进行外膜蛋白(Outer membrane proteins,OMP)分型研究。结果表明:14个禽多杀性巴氏杆菌菌株以2个主要蛋白OmpH和OmpA的差异为依据分为3种主要OMP型;依据次要蛋白的差异,OMP-1,3菌株进一步分为OMP型1.1,1.2和3.1,3.2,其中OMP型1.1有6个菌株,OMP型1.2有5个菌株;OMP型2有1株(YZ7031);P1059为OMP型3.1;CU为OMP型3.2;另外,血清型为A:1的12个菌株中有11个菌株外膜蛋白型均为OMP-1型,血清型为A:3、A:3,4的菌株外膜蛋白型属于3型。说明禽多杀性巴氏杆菌血清型与特定的外膜蛋白型具有很强的相关性。     

Humoral immune responses to Mycoplasma hyopneumoniae in sows and offspring following an outbreak of mycoplasmosis

Previously healthy sows, seropositive to Mycoplasma hyopneumoniae, developed clinical signs of mycoplasmosis, as well as increasing amounts of antibodies to M. hyopneumoniae during an outbreak of the disease in a herd. During the early phase of the outbreak, young piglets (2 weeks) with maternal antibodies remained healthy while older seronegative piglets (4–7 weeks) developed the disease. The duration of the maternal antibodies to M. hyopneumoniae varied between litters and was related to the amount of antibodies in the serum of the dam. In sows, the level of serum antibodies decreased continuously from 4 weeks ante partum to partus, and the level of antibodies in the whey of colostrum was comparable to that in serum 4 weeks ante partum. After loss of maternal antibodies to M. hyopneumoniae, seropositive animals were not found among piglets younger than 9 weeks. Therefore peripheral blood mononuclear cells (PBMC) were collected from various age categories of piglets in order to measure the ability to produce antibodies to M. hyopneumoniae in vitro. PBMC obtained from piglets aged 1 and 3 weeks produced few antibodies to M. hyopneumoniae. Significantly higher levels of antibodies to M. hyopneumoniae were produced by PBMC obtained from pigs aged 5–9 weeks. Thus, the ability of PBMC to produce antibodies to M. hyopneumoniae in vitro seemed to be age-dependent.     

Effect of pneumonia on growth rate and feed efficiency of minimal disease pigs exposed to Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae

Pigs from a minimal disease herd were exposed to pneumonia through contact with pigs infected with Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae. Growth rate, feed efficiency, extent of pneumonial lesions at necropsy and as determined radiographically, and clinical signs of appetite, coughing, sneezing, dyspnea and lethargy were recorded for each pig. Pneumonia occurred as an active, slowly progressive infection during the trial. Coughing was not a good indicator of severity of pneumonia. Increasing severity of pneumonia (measured radiographically or at slaughter) was negatively correlated with performance during the finishing period. Data from this trial support a model that had been developed to relate performance effect to severity of pneumonia.     

SmpB: a novel outer membrane protein present in some Brachyspira hyodysenteriae strains

A novel outer membrane protein-encoding gene was identified in Brachyspira hyodysenteriae. The predicted protein, SmpB, was encoded by a gene that contains regions of identity with that encoding the previously identified lipoprotein SmpA. However, the majority of the reading frame encoding SmpA and SmpB share no detectable similarity. Analysis of several strains revealed that B. hyodysenteriae harbours either smpA or the newly identified gene smpB, but not both. smpB encodes for a slightly larger protein than smpA, 17.6 and 16.8 kDa, respectively. The predicted proteins share an identical leader sequence and the first 10 amino acids of the mature protein, however, the remainder of the predicted protein sequence shows no similarity. It is hypothesised that smpA and smpB are present on the same area of the chromosome. The proteins are antigenically unique, as antisera raised against a strain of B. hyodysenteriae that expresses SmpA cannot detect SmpB and vice versa. Although the presence of an identical leader peptide suggests identical localisation of SmpA and SmpB, it is not known if the two predicted proteins share similar function.     

4株鸭源肠球菌的鉴定和致病性

对临床分离的4株鸭源肠球菌郑1株、郑2株、郑3株、北京株和1株粪肠球菌参考菌株进行了系统鉴定,并用SDS-PAGE和Western-blot技术对各菌株细胞壁蛋白图谱进行比较分析。结果5个菌株的形态、染色、生理生化特性均与粪肠球菌特性一致;它们均对青霉素、万古霉素和庆大霉素敏感而对四环素耐药;5个菌株人工感染雏鸭及小白鼠均有致病性,但各菌株间致病力存在差异,北京株最强,参考株最弱,其余3株介于北京株和参考株之间;各菌株的细胞壁蛋白经SDS-PAGE在相对分子质量33 370~131 690之间均显示数十条蛋白带,其中郑2株和北京株在相对分子质量66 840处均有1条染色较深的蛋白带,而用Western-blot分析显示抗北京株胞壁蛋白抗体只能检测到北京株相对分子质量为66 840的抗原蛋白。以上结果表明,这5个被检菌株为致病性粪肠球菌,且致病性以北京株最强。     

Analysis of the immunoproteome of Mycoplasma mycoides subsp. mycoides small colony type reveals immunogenic homologues to other known virulence traits in related Mycoplasma species

Contagious bovine pleuropneumonia (CBPP) caused by Mycoplasma mycoides subsp. mycoides small colony type (MmmSC) has been eradicated in the developed world, but it is still present in many countries of sub-Saharan Africa. After initially successful control measures in the 1960s it has been spreading due to a lack of money, fragmentation of veterinary services, uncontrolled cattle movement, insufficient vaccine efficacy and sensitivity of current diagnostic tests.In this study we used two-dimensional polyacrylamide gel electrophoresis followed by immunoblot with sera from MmmSC-infected animals and MALDI-ToF mass spectrometry to identify novel immunogenic proteins as candidate molecules for improved diagnostics and vaccines. We identified 24 immunogens recognized by pooled sera from experimentally infected cattle. Furthermore, a serum from an animal with acute clinical disease as well as severe pathomorphological lesions recognized 13 additional immunogens indicating variation in the antibody responses to CBPP amongst cattle. Most immunogens showed compelling similarity to protein/gene sequences in the two ruminant pathogens M. capricolum subsp. capricolum and M. mycoides subsp. mycoides large colony type both belonging to the mycoides cluster. Three of these proteins, namely glycerol-3-phosphate oxidase, adenylosuccinate synthase, and glyceraldehyde-3-phosphate dehydrogenase, had no compelling homologue in the other distantly related bovine pathogen M. agalactiae. In addition, translation elongation factor Tu, heat shock protein 70, pyruvate dehydrogenase, and FKBP-type peptidyl-prolyl isomerase, which have been found to mediate adhesion to host tissue in other mycoplasmas were shown to be expressed and recognized by sera. These proteins have potential for the development of improved diagnostic tests and possibly vaccines.     

In vitro efficacy of a polyhexamethylene biguanide-impregnated gauze dressing against bacteria found in veterinary patients

OBJECTIVE: To evaluate the in vitro efficacy of polyhexamethylene biguanide (PHMB)-impregnated gauze dressing in limiting the growth of bacteria both within and underneath the dressing. STUDY DESIGN: In vitro study. METHODS: Squares of PHMB-impregnated and control gauze were placed on agar plates inoculated with 1 of 11 bacterial species, including 8 multi-resistant organisms. Growth under the gauze was assessed qualitatively after 24-hour incubation. Repeated use of sponges was used to evaluate residual inhibitory activity against Micrococcus lutea and Staphylococcus schleiferi ss. schleiferi. In a second procedure, PHMB-impregnated and control gauze squares were placed in sterile plastic wells and inoculated with 1 of 5 bacterial species, including Pseudomonas spp. and Klebsiella spp. Inhibition of bacterial growth within and underneath the dressing after 24-hour incubation was evaluated by quantifying the numbers of bacteria on the well floor and within each square. RESULTS: PHMB-impregnated gauze provided greater inhibition of growth of 4/4 Gram-positive species and 2/6 Gram-negative species on inoculated plates compared with control gauze. Residual inhibitory activity of PHMB-impregnated gauze was significantly greater against M. lutea on all days and against S. schleiferi ss. schleiferi on days 1 and 4 compared with control. No bacteria were recovered from inoculated PHMB-impregnated gauze squares placed in sterile wells or from the well floor underneath. More than 9 x 10(5) colony-forming units (CFU) were recovered from inoculated control samples placed in sterile wells and more than 8.4 x 10(4) CFU were recovered from control well floors. CONCLUSION: PHMB-impregnated gauze dressing, when placed on inoculated agar plates, reduces growth of underlying bacteria, particularly Gram-positive species. Wet-inoculated PHMB-impregnated dressing prevents growth of Gram-positive and Gram-negative bacteria both within and underneath the dressing. CLINICAL RELEVANCE: PHMB-impregnated dressings may be useful for reducing contamination of underlying wounds by bacterial pathogens.