健康猪肠道菌群的分离培养与耐药性分析

为了解猪肠道正常菌群的耐药性及其临床意义,从健康猪新鲜粪便中,分别采用麦康凯琼脂、MRS增菌液与HYA琼脂、MRS增菌液与TYP琼脂分离大肠杆菌、乳酸杆菌和双歧杆菌,并挑取单个菌落进行形态染色观察、生化鉴定及药敏试验。结果发现,大肠杆菌对克林霉素和红霉素有较强耐药性,乳酸杆菌对诺氟沙星有较强耐药性,而双歧杆菌对诺氟沙星、氯霉素有较强耐药性,提示正常菌群的耐药性可能是影响肠道病原菌感染及其治疗效果的不可忽视因素。     

具有益生特性的小尾寒羊肠道正常菌群的分离、鉴定及筛选

为了获得适合生产的羊源益生菌株,本试验从健康成年小尾寒羊肠道内分离出5株菌落形态各异、生长良好的菌株。通过革兰染色、生理生化鉴定及16SrDNA分析,5株菌分别为M1(干酪乳杆菌)、M2(植物乳杆菌)、E1(粪肠球菌)、E2(屎肠球菌)、L1(枯草芽孢杆菌)。细菌生长曲线、耐酸、耐胆盐、抑菌试验等益生特性分析结果表明,M1、M2和L1具有生长速度快,耐酸、耐胆盐,明显抑制大肠杆菌、鸡白痢沙门菌和金黄色葡萄球菌等肠道常见致病菌的益生特性,可以作为研制羊用微生态制剂的候选菌株。     

Conjunctival bacterial and fungal flora in healthy horses in the UK

Objectives To describe the bacterial and fungal flora of the normal conjunctiva of horses in the UK; to determine the effect of horse age, sex, geographic location, and housing on this flora; and to determine the most appropriate antimicrobial drug(s) for prophylactic treatment of corneal ulcers. Animal studied A total of 60 adult healthy horses were studied. Procedure Swabs of the conjunctiva were obtained from 60 horses housed in two locations within the UK. Specimens were cultured for aerobic bacteria and fungi, and sensitivity against six ophthalmic antimicrobials assessed. The effect of age, sex, location and housing on the frequency of microbial isolation was evaluated. Results Fifty‐four bacterial isolates, representing thirteen genera of bacteria were cultured from 31 (52%) horses. The most frequently isolated bacterial species was Acinetobacter sp (17/32 horses). The majority of isolates (28/54: 52%) were gram‐positive. Three genera of fungi (Mucor, Absidia and Aspergillus spp) were isolated from eight (13%) horses. There was no significant effect of geographic location, sex, age or housing on frequency of microbial isolation. Horses from which gram‐negative bacteria were isolated were significantly older than horses from which gram‐positive bacteria were isolated. High efficacy (greater than 90% of isolates sensitive in vitro) was displayed by chloramphenicol, gentamicin and tetracycline. Conclusions The microbial species isolated are comparable with studies performed in other countries, although the frequency of Acinetobacter isolation was higher which may reflect a geographic difference. The topical antimicrobials gentamicin and chloramphenicol are appropriate first line antimicrobials for empirical treatment of corneal ulcers in the UK.     

Isolation and distribution of bacterial flora in farmed rainbow trout from Mexico

Trout farming is a growing aquaculture industry in Mexico, with stock mainly supplied by the importation of eyed eggs. The aim of the present study was to determine the frequency of bacterial isolations in farmed rainbow trout Oncorhynchus mykiss from Mexico. Sixty-five farms distributed among seven states of Mexico were included in the study. Individual samples from gills, liver, spleen, intestine, and kidney were obtained from 563 apparently healthy fish. In total, 371 bacterial isolates were recovered from sampled fish; isolates of the genera Aeromonas, Edwardsiella, Enterobacter, Escherichia, Klebsiella, Plesiomonas, Pseudomonas, and Yersinia were identified. Aeromonads were the most frequently isolated bacteria. Renibacterium salmoninarum was not isolated from any of the sampled fish. Our results showed the presence of bacteria that are potential pathogens of both rainbow trout and humans.     

Staphylococcus sciuri as a part of skin, nasal and oral flora in healthy dogs.

The coagulase-negative species Staphylococcus sciuri is widespread in nature and is associated with a variety of domestic and wild animals. However, the occurrence of S. sciuri in dogs has received little attention so far. In the present study, we established the prevalence of S. sciuri in a large population of healthy dogs, and characterized isolated strains. Samples from two mucous membrane sites (anterior nares and mouth), and two hair-coated sites (head and withers) were taken from 122 dogs and inoculated into STS agar, a novel selective medium that was introduced and tested in the study. In total, 116 isolates of S. sciuri were obtained from 488 specimens. S. sciuri was isolated from 56 out of 122 (46%) dogs. The occurrence of S. sciuri in the anterior nares and mouth were significantly higher than those in withers and head. No significant association of S. sciuri occurrence in dogs and factors such as sex, age, and living environment (indoor/outdoor) was found. Out of 56 dogs, which tested positive for S. sciuri, 30 (54%) would have it as a resident flora. Thus, we showed that S. sciuri was frequently present as a part of skin, nasal and oral flora in healthy dogs both as a resident and transient carriage.     

Isolation of Actinomyces hyovaginalis from sheep and comparison with isolates obtained from pigs

Actinomyces hyovaginalis, an organism initially described from pigs, was recovered from nine sheep and a moufflon. Further strains of A. hyovaginalis were recovered from five samples from pigs over the same period. 16S rRNA sequencing and extensive phenotyping demonstrated high similarity between the ovine and porcine isolates; however differences with respect to erythritol, adonitol and l-arabitol fermentation were detected. Ovine isolates were made from various sample sites including abscesses and highlight the importance of the accurate identification of the various coryneform isolates which affect sheep. A. hyovaginalis can be added to the growing list of coryneforms which can cause disease in sheep including Corynebacterium pseudotuberculosis, Trueperella pyogenes and Arcanobacterium pluranimalium.     

Comparison of the bacterial flora of the duodenum in healthy cats and cats with signs of gastrointestinal tract disease

OBJECTIVE: To determine whether a colony environment predisposes healthy cats to high bacterial counts, including counts of obligate anaerobes, in the duodenum and whether increased numbers of bacteria could be found in the duodenum of cats with signs of chronic gastrointestinal tract disease. DESIGN: Prospective study. ANIMALS: 20 healthy control cats (10 from a colony environment and 10 pet cats) and 19 cats with a history of chronic gastrointestinal tract disease. PROCEDURE: Undiluted duodenal fluid was quantitatively and qualitatively assessed by bacteriologic culture under aerobic and anaerobic conditions. Serum concentrations of cobalamin and folate were also measured. RESULTS: Significant differences were not detected in the numbers of bacteria found in the duodenum of cats housed in a colony environment, compared with pet cats fed an identical diet prior to sampling. All healthy cats were, therefore, combined into 1 control group. Compared with healthy cats, cats with clinical signs of gastrointestinal tract disease had significantly lower counts of microaerophilic bacteria, whereas total, anaerobic, and aerobic bacterial counts were not significantly different. None of the cats with disease had total bacterial counts higher than expected from the range established in the control cats. Differences were not detected in regard to serum folate or cobalamin concentrations between diseased and healthy cats. CONCLUSIONS AND CLINICAL RELEVANCE: These findings indicated that healthy colony cats and pet cats have high numbers of bacteria in the duodenum, including high numbers of obligate anaerobes. Our findings also suggest that bacterial overgrowth in the small intestine is not a common clinical syndrome in cats with chronic nonobstructive gastrointestinal tract disease.     

健康与子宫内膜炎奶牛阴道菌群结构的比较研究

利用16SrDNA结合PCR-DGGE技术分析比较了产后45d健康奶牛和患子宫内膜炎奶牛阴道内细菌群落多样性及其差异,并通过切胶、克隆测序等步骤分析了奶牛阴道细菌种类。研究结果表明,患病奶牛DGGE图谱的条带数量、多样性指数均显著高于健康奶牛;健康奶牛阴道优势菌主要有清酒乳杆菌(Lactobacillus sakei subsp.)、魏斯杆菌(Weissella koreensis sp.)和地衣芽孢杆菌(Bacillus licheniformis),而患病组奶牛阴道内此3种优势菌含量显著降低或缺失,且也无其他明显优势菌存在;从健康和患病奶牛阴道内均分离出脆弱拟杆菌(Bacteroides fragilis)、溃疡拟杆菌(Bacteroides helcogenes)、无乳支原体(Mycoplasma agalactiae)、普氏菌(Prevotella dentalis)、产气荚膜梭菌(Clostridium perfringens str.)和肠球菌(Enterococcus hirae)等常见病原微生物,在患病奶牛阴道内分离出大肠杆菌(Escherichia coli str.K-12 substr.)、梭杆菌(Fusobacterium nucleatum subsp.)等特定致病菌。研究结果提示,子宫内膜炎奶牛阴道内菌群结构比健康奶牛更为复杂,且无优势菌,阴道菌群结构失衡可能是奶牛发生子宫内膜炎的原因。     

Flubendazole metabolism and biotransformation enzymes activities in healthy sheep and sheep with haemonchosis

Bártíková, H., Křížová, V., Lamka, J., Kubíček, V., Skálová, L., Szotáková, B. Flubendazole metabolism and biotransformation enzymes activities in healthy sheep and sheep with haemonchosis. J. vet. Pharmacol. Therap . 33 , 56–62.
The aim of this project was to study the influence of haemonchosis, a common parasitic infection of small ruminants caused by Haemonchus contortus , on the activity of biotransformation enzymes and on in vitro flubendazole (FLU) biotransformation in liver and small intestine of lambs ( Ovis aries ). Twelve lambs were divided into three groups: non-infected animals, animals orally infected with larvae of H. contortus ISE strain for 7 weeks and for 11 weeks. At the end of the experiment, hepatic and intestinal subcellular fractions were prepared and used for assays of biotransformation enzymes activities and FLU metabolism testing. The activities of hepatic cytochromes P450, flavine monooxygenases and carbonyl-reducing enzymes were decreased in infected animals. UDP-glucuronosyl transferase activity was significantly lower (by 35%) in 11 weeks infected animals than that in control animals. When in vitro metabolism of FLU was compared in control and infected animals, significantly lower velocity of FLU reduction was found in infected animals. Slower FLU reduction may be beneficial for the haemonchosis treatment using FLU, because FLU will remain longer in the organism and could cause longer contact of parasites with FLU.     

A comparison of corneal sensitivity between healthy cats and cats with corneal sequestra

In order to establish reference values for corneal sensitivity in ophthalmologically healthy persians (n = 40) and domestic short hair cats (n = 60) a prospective study was conducted. Furthermore corneal sensitivity in 48 cats with a corneal sequestrum was measured. Corneal sensitivity was recorded with the help of the aesthesiometer according to Cochet and Bonnet in five different corneal locations (central, nasal, dorsal, temporal, and ventral). The sensitivity for the central corneal region was recorded as amounting to 3.58 +/- 0.56 cm in ophthalmologically healthy domestic short hair cats and to 2.97 +/- 0.58 cm in healthy persian cats. The sensitivity of the central corneal area of a cat with a corneal sequester only amounts to 2.03 +/- 0.53 cm. Between the diseased and the healthy eyes no statistical difference could be demonstrated for any of the measured corneal locations. The sensitivity of the peripheral corneal locations is significantly lower than that of the central corneal region in all three groups examined.     

Teat skin normal flora and colonization with mastitis pathogen inhibitors

Isolates of bacteria from normal teats were used to attempt colonization of teats of dry cows or neonatal calves. Isolates for inoculation were chosen on the basis of ability to inhibit mastitis pathogens in vitro, with the ultimate goal of in vivo inhibition of mastitis pathogens at the teat surface. Three bacterial normal flora isolates (Corynebacterium xerosis, Bacillus sp. and Aerococcus viridans) persisted less than 10 days on the teats of dry cows. The fourth isolate, Staphylococcus hominis 1, was studied in greatest detail because studies characterizing the normal teat flora showed staphylococci to be the predominant flora. The S. hominis 1 isolated used for inoculation was an inhibitor of Gram-positive mastitis pathogens. It was a biotype not found on these teats prior to inoculation, thus facilitating identification of the inoculated isolate on sequential sampling. Colonization of newborn calves, before other bacterial floral became established, resulted in recovery of inoculated S. hominis 1 for an average of 51 days or longer. On dry cow teats it was detected for up to 28 days. On several occasions the inoculated S. hominis 1 was found in pure culture. Since many new infections occur during the dry period, the colonization of dry cow tests with S. hominis 1 organisms inhibitory for Gram-positive pathogens should be tested as an adjunct to other methods of mastitis prevention.     

Intestinal bacterial flora of the household lizard, Gecko gecko

A total of 114 isolates was recovered from the intestines of 43 househould lizards, Gecko gecko. Among the important ones were Staphylococcus aureus, Salmonella typhimurium, Pseudomonas aeruginosa, Proteus mirabilis and Edwardsiella tarda.     

Cloning and comparison of bighorn sheep CD18 with that of domestic sheep, goats, cattle, humans and mice

Previously, we have shown that CD18, the beta-subunit of beta(2)-integrins, serves as a receptor for leukotoxin (Lkt) secreted by Mannheimia (Pasteurella) haemolytica on bovine leukocytes. Anti-CD18 monoclonal antibodies (mAbs) inhibit Lkt-induced cytolysis of bighorn sheep (Ovis canadensis) leukocytes suggesting that CD18 may serve as a receptor for Lkt on the leukocytes of this species as well. Confirmation of bighorn sheep CD18 as a receptor for Lkt, and elucidation of the enhanced Lkt-susceptibility of bighorn sheep polymorphonuclear leukocytes (PMNs), necessitates the cloning and sequencing of cDNA encoding bighorn sheep CD18. Hence, in this study we cloned and sequenced the cDNA encoding CD18 of bighorn sheep, and compared with that of other animal species. The cDNA of bighorn sheep CD18 has an open reading frame (ORF) of 2310bp. CD18 sequences obtained individually from peripheral blood mononuclear cells (PBMCs) and PMNs were identical to each other. Comparison of the deduced 770-amino acid sequence of CD18 of bighorn sheep with that of domestic sheep, goats, cattle, humans and mice revealed 99, 98, 95, 82 and 80% identity, respectively. Availability of cloned bighorn sheep CD18 cDNA should allow the molecular characterization of M. haemolytica Lkt-receptor interactions in bighorn sheep and other ruminants that are susceptible to this disease.     

Evaluation of arthrocentesis site bacterial flora before and after 4 methods of preparation in horses with and without evidence of skin contamination

OBJECTIVE: To evaluate the effectiveness of four methods of povidone-iodine preparation on skin bacterial flora of arthrocentesis sites, in horses, with and without evidence of skin contamination. STUDY DESIGN: Prospective randomized study. ANIMALS: Twenty-four adult horses. METHODS: Horses were assigned to either the clean or contaminated group based on housing environment and visual evidence of contamination. Using a moist sterile swab, microbial culture samples were obtained from the skin over the distal interphalangeal joints immediately before and after preparation. Each site was aseptically prepared with 1 of 4 povidone-iodine techniques: 10-minutes scrub, 5-minutes scrub, three 30-second scrubs, or commercial one-step iodophor surgical solution. Colony forming units (CFUs) were determined for each sample, 24 hours after inoculation, on blood agar plates. RESULTS: Mean (+/-SD) pre-scrub CFUs/mL was significantly higher in the contaminated group (9588.33+/-1223.65) compared with the clean group (4489.00+/-3842.03) (P<.01). After preparation of the arthrocentesis sites, there were no significant differences in post-scrub CFUs/mL among the 10 minutes (mean clean, 46.00+/-64.36; mean contaminated, 28.67+/-18.04), 5 minutes (mean clean, 84.17+/-109.80; mean contaminated, 40.33+/-44.52), three 30 seconds povidone-iodine scrubs (mean clean, 95.50+/-172.29; mean contaminated, 46.67+/-56.94), or application of a commercial one-step iodophor surgical solution (mean clean, 102.17+/-161.78; mean contaminated 117.67+/-143.78); or between the clean (81.96+/-131.69) and contaminated groups (58.33+/-85.90) (P<.01). CONCLUSIONS: Preparation of the distal interphalangeal joint arthrocentesis site with each of these techniques significantly reduces the bacterial flora to a similar level for arthrocentesis in horses with and without evidence of skin contamination. Clinical Relevance- Aseptic preparation of the skin over the distal interphalangeal joint may be accomplished with any of these techniques.