Induction of Escherichia coli mastitis in cows fed selenium-deficient or selenium-supplemented diets

Ten Holstein heifers were fed a selenium-deficient (SeD) diet (0.04 mg of Se/kg on a total ration dry-matter basis) 3 months before calving and throughout their first lactation. A selenium-supplemented (SeS) diet (2 mg of Se/head/d) was fed to a group of 10 heifers. In about the 14th week of lactation, the cows were challenge-exposed to Escherichia coli by administering 15 to 40 colony-forming units (CFU) into 1 mammary gland. Selenium concentration (microgram/ml) in blood around the time of challenge exposure was 0.033 +/- 0.002 (mean +/- SEM) in SeD and 0.132 +/- 0.006 in SeS cows. Infections were established in all challenge-exposed quarters. The frequency of quarter atrophy and agalactia, and reduction in whole-udder milk yield in the first 4 days after challenge exposure, were greater (P less than 0.05) in the SeD cows. Log10 peak bacterial concentrations in milk were higher (P less than 0.05) in SeD (7.63 +/- 0.34 CFU/ml) than in SeS cows (5.57 +/- 0.66 CFU/ml). Mean log bacterial concentration was significantly higher (P less than 0.05) from 12 to 20 hours after challenge exposure in SeD than in SeS cows. Duration of infection was significantly greater (P less than 0.05) in SeD (162.0 +/- 12.0) than in SeS cows (114.4 +/- 18.0 hours). Milk somatic cell counts increased significantly more slowly (P less than 0.05) in SeD than in SeS cows from 8 to 16 hours after challenge exposure. Ratios of milk somatic cells to bacteria in milk were significantly lower (P less than 0.05) in SeD than in SeS cows at 12 and 16 hours after challenge exposure.     

Waterlogging tolerance in lentil (<Emphasis Type="Italic">Lens culinaris</Emphasis> Medik. subsp. <Emphasis Type="Italic">culinaris</Emphasis>) germplasm associated with geographic origin

Variation in response to climatic and edaphic factors has resulted in the geographic differentiation and specific adaptation of lentil germplasm to its environment. The germination of lentil seeds can be affected by waterlogging caused by relay sowing into standing rice crops. Consequently, there is a demand for waterlogging-adapted lentil genotypes, particularly at germination. This experimentation investigated the effects of waterlogging at germination on a germplasm collection. Firstly, two contrasting genotypes, ATC 70876 and Nugget, were waterlogged for six durations (0, 3, 6, 9, 14 and 21 days) in a pot soil system in the glasshouse. Six days of waterlogging was the duration for germination where genotypes showed the largest contrast in germination rate; therefore, this duration was applied to a larger number (127) of genotypes from 11 countries in a screening trial. Finally, all 127 genotypes were grown in pots in the open to phenotype morphological traits (seed size, above-ground biomass, time to flower and maturity). There was significant variation among genotypes, and those originating from Bangladesh had the highest germination at 21.2 %. The heritability of germination on waterlogged soil among genotypes was intermediate at H² = 44.8 %. Correlations between morphological and physiological traits showed that the capability of seeds to germinate on waterlogged soil was negatively correlated to time to maturity (r = ?0.643). It was concluded that genotypes from Bangladesh are adapted to waterlogging at germination. With an intermediate heritability for germination under waterlogging conditions, selection for relay sowing through waterlogging tolerance at germination is practicable.     

Growth effects in tropical nickel‐agromining ‘metal crops' in response to nutrient dosing

Agromining is an emerging technology that utilizes selected ‘metal crops' (= hyperaccumulator plants) to extract valuable target metals from unconventional resources for profit from mineralised soils. Growth characteristics, shoot metal concentrations, and agrominable locations are important considerations in economic agromining. Globally, the greatest potential for nickel (Ni) agromining exists in the tropics. However, the agronomic systems of tropical ‘metal crops' have not been previously tested. Furthermore, it is currently unknown whether nutrient dosing of prospective tropical agromining Ni ‘metal crops’ could possibly cause a shoot Ni‐dilution effect which may ultimately limit economically viable Ni yields. We undertook a pioneering study on Ni uptake and growth responses to nutrient dosing in two promising tropical ‘metal crops' (Phyllanthus rufuschaneyi and Rinorea cf. bengalensis). The experiment consisted of a large randomised block growth trial in large pots over 12 months in Sabah (Malaysia). At 3‐month intervals, the plants were exposed to soluble treatments that altered available concentrations of nitrogen (N), phosphorus (P), and potassium (K). We found strong positive growth responses to N and P additions in P. rufuschaneyi, whereas K additions had negative growth effects. In R. cf. bengalensis, all treatments had positive growth effects. The increases in biomass in response to nutrient dosing did not significantly reduce shoot Ni concentrations in both species, with the exception of N addition in P. rufuschaneyi. This study reveals that Ni uptake and growth responses to nutrient dosing are species‐dependent, primarily influenced by the ecophysiology of the species. Inorganic fertilization could possibly be an important component of the management of local ‘metal crops' to be used in viable commercial agromining in the tropics, but this needs to be tested in the field with different formulations of N, P, and K.     

Molecular analyses of mycobacteria other than the M. tuberculosis complex isolated from Northern Ireland cattle

Mycobacteria other than the Mycobacterium tuberculosis complex (MOTT), isolated from Northern Ireland cattle, were identified by PCR amplification of the 16S rRNA gene, and subsequent reverse cross blot hybridisation and sequence analyses. Elucidation of the MOTT species was to facilitate specificity testing of new and existing diagnostic test reagents for bovine tuberculosis. The presence of the genes for potential diagnostic antigens: MPB70, MPB64, ESAT-6 and CFP-10 in the isolated MOTT species was investigated. Molecular analyses of cultured isolates from bovine lymph node specimens of 48 cattle identified a wide variety of mycobacterial species including Mycobacterium nonchromogenicum, Mycobacterium malmoense, Mycobacterium bohemicum, Mycobacterium paratuberculosis, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium holsaticum, Mycobacterium palustre, Mycobacterium sp. IWGMT 90210, Mycobacterium sp. LIV-2129, a potentially novel mycobacterial species (EMBL/GenBank/DDBJ Accession Number AJ617495) and Rhodococcus equi. Apart from M. kansasii, the results of traditional (standard phenotypic and biochemical) and molecular identification methods did not correlate well, with traditional methods identifying fewer species. Most of the species identified were either recognised pathogenic or potential pathogenic species. The genes for ESAT-6, CFP-10 and, unusually, MPB64 were detected in M. kansasii only. The MPB70 gene was not detected in any of the species. This study supported restricted species distribution of these genes as well as identifying a different range of MOTT species that could be included in specificity testing of new diagnostic reagents for bovine tuberculosis.     

Pseudomonas mastitis: difficulties in detection and elimination from contaminated wash-water systems

Histories of 4 dairy herds with increased incidence of Pseudomonas mastitis associated with contaminated wash hoses in milking parlors are described. Problems of detection and elimination of the organism from contaminated water sources and the inadequacy of iodide germicides in eliminating Pseudomonas are discussed. In problem herds, greater numbers of organisms often are found in the water left standing in the wash hoses between milkings. Thus, flushing of hoses before their use in the milking process may be beneficial in reducing exposure of the cows to the organism.     

Post-milking teat dip use in dairy herds with high or low somatic cell counts.

Milk samples for bacteriologic culture were submitted from 71 dairy herds, 24 with low somatic cell count (SCC) and 47 with high SCC and high prevalence of subclinical mastitis. At the time of sample submission to the Mastitis Diagnostic Laboratory of Pennsylvania State University, information regarding the herd mastitis control practices was collected. A combined program of post-milking teat dipping (PMTD) and antibiotic treatment of all cows at the start of the nonlactating period was practiced more frequently for herds with low SCC, (P less than 0.001) than for herds with high SCC. Among all herds for which PMTD was practiced, a higher proportion (P less than 0.001) of those for which chlorhexidine-based products were used had low SCC than high SCC. Conversely, a higher proportion of herds for which a dip with an acrylic latex barrier was used had high SCC rather than low SCC (P = 0.002). For herds with high prevalence of subclinical mastitis, and despite a program of PMTD and treatment of all cows at the start of the nonlactating period, a change to a different germicidal teat dip product may be indicated to help reduce prevalence of infection.     

Intramammary administration of gentamicin as treatment for experimentally induced Escherichia coli mastitis in cows.

In 8 Holstein cows, 50 colony-forming units (CFU) of Escherichia coli was administered into 1 mammary gland. Infections were established in all inoculated glands. In 4 of the 8 cows, 500 mg of gentamicin sulfate was administered by intramammary infusion 14 hours after inoculation; the other 4 cows were untreated controls. Infusions of gentamicin also were given after each of the 3 successive milkings after the initial infusion, so that a total dose of 2 g of gentamicin was given to each of the treated cows. During the 33-hour treatment period and for the first milking after the last infusion of gentamicin, the treated cows had a mean gentamicin concentration of greater than or equal to 31.0 micrograms/ml in milk samples that were collected from inoculated quarters immediately before each milking. Concentrations of 0.34 and 0.69 micrograms of gentamicin/ml were detected in milk from 2 cows at 8 days after inoculation with E coli. Mean serum concentrations of gentamicin were greater than or equal to 0.37 micrograms/ml throughout the treatment period and the first 12 hours after the last infusion, with a mean peak concentration of 0.96 micrograms/ml at 24.4 hours. The range of peak concentration of gentamicin detected in urine from all treated cows was 42 to 74.4 micrograms/ml. Peak concentration of E coli in milk in the treated cows (6.08 +/- 1.02 log10 CFU/ml) did not significantly (P greater than 0.05) differ from that of the control cows (5.26 +/- 1.00 log10 CFU/ml). Similarly, mean duration of infection in the treated cows (54 hours) did not differ significantly from that of the control cows (48 hours).(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation and genotyping of Campylobacter species from kiwi (Apteryx spp.) in captivity: implications for transmission to and from humans

Aims: To investigate the presence of Campylobacter spp. in captive kiwi (Apteryx spp.) and compare their genotypic profiles with those of human and animal origin, in order to assess their potential for zoonotic or zooanthroponotic transmission.

Methods: Conventional selective enrichment and filter-based isolation methods were applied to isolate Campylobacter spp. from fresh faecal samples from 12 North Island brown kiwi (Apteryx mantelli) and one great spotted kiwi (A. haastii), housed in one of five different areas in a kiwi sanctuary in Christchurch, New Zealand. Isolates were identified using multiplex PCR and 16S rRNA gene sequencing. High-resolution rapid genotyping using multiplex ligation-dependant probe amplification-based binary typing (MBiT) was applied and profiles compared with similar results from 2,165 Campylobacter spp. isolates contained in a database derived from human clinical, veterinary and environmental samples.

Results: One isolate of C. jejuni, and one belonging to the C. lari phylogenetic group were recovered from faeces from two kiwi. High-resolution rapid genotyping by MBiT demonstrated these to be indistinguishable from isolates obtained previously from human cases of diarrhoea, and others from chicken, cattle, sheep and water.

Conclusions: These data provide evidence for potential zoonotic or zooanthroponotic transmission of Campylobacter spp. in kiwi with implications for management of birds kept in captivity. We believe this is the first formal report of C. jejuni and a C. lari-like organism in kiwi.

Abbreviations: MBiT: Multiplex ligation-dependant probe amplification-based binary typing