Bacterial and fungal pathogens isolated from corneal ulcerations in domesticated elephants (Elephas maximus maximus) in Sri Lanka

Infective corneal diseases are common among the domesticated elephants in Sri Lanka, causing morbidity including blindness leading to loss of revenue to their owners. From 140 elephants of different ages and both sexes, 36 animals (25.7%) had evidence of keratitis, corneal ulcers, corneal opacities, and some had foreign bodies in their eyes. Nine elephants (6.4%) had lesions in both eyes (6.41%). Cultures for both bacteria and fungi were obtained from 26 corneal ulcers, including the nine elephants with bilateral lesions. The other 10 animals could not be restrained for sample collection. Swabs from the normal corneas of an additional 20 elephants without signs of any ophthalmic diseases were also collected. Twenty-three of the 35 (65.71%) samples from affected corneas yielded bacterial pathogens, and 14 (40%) also had fungal isolates. None of them yielded a fungal isolate alone. The predominant bacteria isolated were Staphylococcus aureus , beta hemolytic streptococci and coliforms. Fusarium , Cladosporium , Curvularia and Aspergillus species were the primary fungal isolates. No bacteria or filamentous fungi were isolated from the eyes with the normal corneas. Microbial identification including fungal isolations is suggested in the management of infective corneal diseases in elephants.     

Impact of Mastitis Control Measures on Milk Production and Mastitis Indicators in Smallholder Dairy Farms in Kiambu District,Kenya

Bovine mastitis and mastitis control were investigated on smallholder farms in central Kenya. After an initial observational study, a clinical trial to assess the impact of three different mastitis control strategies – (1) improved udder hygiene, (2) treatment of subclinical cases, and (3) a combination of these – was conducted on 100 randomly selected farms with 332 lactating cows. Before the implementation of control measures, the milk yield was low (mean 6.5 kg/day; median 6 kg/day) and somatic cell counts (SCC) were high, with 80% and 43% of cows having milk with SCC greater than 250×10³ cells/ml and 600×10³ cells/ml, respectively. Infectious pathogens were also commonly isolated, with 63% of cows being positive for pathogenic bacteria. Neither intervention strategy alone had any effect on mastitis indicators or milk yield. In combination, the measures had some impact, lowering the prevalence of contagious pathogens by 18%, but this was not reflected in a significantly increased milk yield, lowered SCC or reduced incidence of clinical mastitis.     

Mastitis Whey — a Good Medium for Bacteria?

Growth of mastitis pathogenic bacteria was measured in bovine whey samples by a turbidometric microtechnique. Whey from mastitis cows supported growth as compared with whey prepared from normal milk. Blood proteins leak into milk during mastitis. A study was undertaken to analyze which molecules from blood would promote bacterial growth in whey Fractions containing hemoglobin showed a distinct growth-promoting effect. An inadequate iron supply is one of the restricting growth factors for bacteria in milk. By utilizing heme-compounds the pathogens can by-pass the effect of antimicrobial iron-binding present in milk in the form of lactoferrin.     

Seasonal effects on the aerobic bacterial and fungal conjunctival flora of normal thoroughbred brood mares in Florida

Objective To evaluate seasonal effects on the presence or absence of fungal and aerobic bacterial flora of the conjunctival fornix of normal Florida Thoroughbred horses. Sample population Both eyes of 100 horses. Procedure Horses with normal anterior segment ophthalmic examinations from three farms in north central Florida were included. Each animal had the ventral conjunctival fornix of each eye swabbed with sterile culturettes. Samples were taken in October, January, April, and July (1999–2000). Aerobic and fungal cultures were plated. Bacterial cultures were reviewed at 24 and 48 h. Fungal cultures were reviewed weekly for 4 weeks. Logistic regression analysis with season as a factor and age of the horse as a covariate was performed. Statistical significance was set at P < 0.01. Results Horses ranged from 3 to 24 years of age, with a median age of 9 years. Twenty‐four genera of bacteria and 35 genera of fungi were recovered. Corynebacterium sp., Staphylococcus sp., Bacillus sp. and Moraxella sp. were the bacteria most frequently isolated. Mold species, dematiaceous mold species, Chrysosporium sp., Cladosporium sp., and Aspergillus sp. were the most frequently recovered fungi. Season did not have a significant effect on the presence of microorganisms isolated for individual horses adjusted for age. Younger horses had an increased incidence of gram‐negative rods and fungal isolates. The number of bacteria and fungi isolated are not uniform across seasons. Conclusion There were no significant differences between the number or type of organisms cultured during the sampling seasons in normal Florida horses. A large range of normal bacterial and fungal flora were isolated from these horses. The number of bacteria and fungi isolated are not uniform across seasons. The likelihood of detecting an organism depends on the horses’ age.     

Prevalence and Aetiology of Mastitis in Cows from Two Major Ethiopian Dairies

The study was undertaken to determine the aetiology and prevalence of mastitis in hand-milked cows (n = 186) in two major Ethiopian dairies. The California Mastitis Test and culturing for bacteria revealed that 21.5% of the cows were clinically infected and 38.2% had subclinical mastitis. Most mastitis pathogens isolated from milk samples testing positive by the California Mastitis Test were Gram-positive cocci. Staphylococci constituted 57% of the isolates, of which the predominant cause of bovine mastitis was Staphylococcus aureus (40.5%). Other mastitis pathogens isolated include streptococci (16.5%), coliforms (9%) and corynebacteria (5%). Retrospective analysis of farm records indicated that mastitis was the second most important cause of culling and accounted for 27% of the cows removed from these two dairies.     

Protective Effects Against Erwinia amylovora Induced by hrp Mutants in the Whole Plant are Only Partially Mimicked in Cultivated Cells

A virulent strain of Erwinia amylovora, the causal agent of fire blight of Maloideae, and two of its non-virulent hrp mutants (a secretory and a regulatory mutant) were inoculated into apple cell suspensions either alone or in mixed inoculations. In single inoculations, death of 4- to 5-day-old apple cells occurred only when the concentration of the virulent strain of E. amylovora reached a threshold inoculum concentration of 10⁴CFUml^–1, while high concentrations of the hrp mutants were unable to kill apple cells. When mixed inoculated with the virulent parental strain, both hrp mutants protected apple cells from death caused by the virulent strain. The protective effect was associated with a decrease in the population level of the virulent strain and it was dependent on the non-virulent to virulent inoculum concentration suggesting a competition between the non-virulent mutant and the virulent strain. However, no differential protective ability between the two types of mutants could be noticed, contrary to previous results obtained with apple seedlings or apple flowers in which the regulatory mutant was significantly more effective than the secretory mutant. In conclusion, inoculation of apple cell cultures with E. amylovora does not seem to be a model suitable for investigating mechanisms leading to protection.     

Bacterial infection of the lower respiratory tract in 34 horses

OBJECTIVE: To investigate associations between the bacteriology and aspects of history, clinical presentation, outcome and pathology of lower respiratory tract disease of 34 horses. PROCEDURE: Detailed aerobic and anaerobic bacteriological investigations were performed on clinical specimens from horses with pneumonia, lung abscessation and necrotic pneumonia with or without pleurisy in an attempt to identify those bacteria that might contribute to the initiation and progression of infection. RESULTS: Bacteria were cultured from 33 of the 34 horses. In ten cases, only aerobic/facultatively anaerobic isolates were cultured while aerobic/facultatively anaerobic bacteria and obligately anaerobic bacteria were isolated in the other 23 cases. Moderate to large numbers of anaerobic bacteria were isolated only when the estimated duration of illness was at least five days. Bacteria were not cultured from 12 of the pleural fluid samples but were always cultured from pulmonary samples (either transtracheal aspirates from live horses or pulmonary lesions at necropsy). Streptococcus equi subsp zooepidemicus was isolated in the three cases where only one bacterial species was cultured. In the other 30 cases, multiple species were isolated. These included most often and in greatest numbers, Streptococcus equi subsp zooepidemicus, Pasteurellaceae, Escherichia coli, anaerobic cocci, Eubacterium fossor, Bacteroides tectum, Prevotella heparinolytica, Fusobacterium spp, and pigmented members of the genera Prevotella and Porphyromonas. Aerobic/facultatively anaerobic organisms were isolated from 97% of horses, while obligately anaerobic organisms were cultured from 68% of horses. CONCLUSION: There was no association between the isolation of any specific bacterium and the outcome of disease. However, obligately anaerobic bacteria (such as anaerobic cocci, Bacteroides tectum, P heparinolytica and Fusobacterium spp) and the facultatively anaerobic species Escherichia coli, were recovered more commonly from horses that died or were euthanased than from those that survived. There was an association between failure of horses to recover from pleuropneumonia and delay in diagnosis and initiation of treatment.     

Efficacy of Chlorhexidine against Some Strains of Cultured and Clinically Isolated Microorganisms

The efficacy of chlorhexidine digluconate was determined against some strains of collected and clinically isolated bacteria and fungi. The efficacy was evaluated either by calculating a minimum inhibitory concentration (MIC) or by efficacy trials according to the guidelines of the European Committee for Standardization. The MIC values of chlorhexidine for Staphylococcus aureus, Microsporum gypseum, Microsporum canis and Trichophyton mentagrophytes were 0.625 g/ml, 12.5 g/ml, 50 g/ml and 6.25 g/ml, respectively. The in vitro efficacy of chlorhexidine was higher against ATCC strains of S. aureus and P. aeruginosa (0.5 mg/ml for 5 min and 0.5 mg/ml for 10 min, respectively) than against clinical isolates (0.5 mg/ml for 15 min and 1 mg/ml for 10 min, respectively). The antiseptic activity of aqueous solutions of chlorhexidine against spores of Bacillus subtilis, Bacillis sfericus and Clostridium perfringens required longer contact times than against the vegetative forms. Nevertheless, 5 mg/ml of chlorhexidine in water–ethanol 20:80 v/v was totally effective against the vegetative forms or spores of these microorganisms.     

乳酸菌发酵青贮对奶牛产奶量及乳成分的影响

利用从优质青贮饲料中分离鉴定并筛选的2株乳酸菌进行全株玉米的青贮中试试验,玉米青贮30天即可成熟。感官评定的结果表明,乳酸菌处理组的青贮饲料品质比对照组显著提高(p<0.05)饲喂添加乳酸菌的玉米青贮饲料对奶牛的产奶量没有显著影响,但是牛乳中蛋白质和非脂乳固体的含量也显著提高(p<0.05),而对乳脂肪的含量没有影响。     

Validation and application of real-time polymerase chain reaction assays for representative rumen bacteria

Real‐time polymerase chain reaction (PCR) assays for 11 representative rumen bacterial species were validated. The sensitivity was tested by using the serially diluted target 16S rDNA from respective bacterial species. The recovery of the target DNA and the assay reproducibility were determined using DNA from rumen fluid spiked with different quantities of the target. Minimum detection levels for the target were 10–100 copies in pure culture. The recovery of the added target ranged from 82.4 to 116.6%. The intra‐ and inter‐assay variations of each assay were <9.4 and <12.6%, respectively. Therefore, the real‐time PCR assays evaluated in the present study are considered to be sufficiently reliable for monitoring all 11 bacterial species in the rumen. The assays were then applied to the monitoring of the bacterial species attached to ruminally incubated rice straw. Among the monitored fibrolytic species, Fibrobacter succinogenes was found to be the most dominant, accounting for 2.61% of total bacteria after 24 h incubation. Selenomonas ruminantium and Streptococcus bovis, non‐fibrolytics, were detected on the rice straw at 8.96% and 1.16% of total bacteria, respectively. Such high levels of non‐fibrolytics on the plant fiber suggest a synergistic relationship between fibrolytics and non‐fibrolytics.