A Severe Hellenic CMV Tomato Isolate: Symptom Variability in Tobacco, Characterization and Discrimination of Variants

A severe strain of Cucumber mosaic virus (CMV) originating from an infected tomato plant (Gastouni-Olympia, Greece) was isolated in tobacco (Nicotiana tabacum cv. Xanthi nc), after three serial local lesion passages in Chenopodium quinoa and designated CMV-G. CMV-G induces yellow mosaic (YM) symptoms in tobacco. When CMV-G was passed mechanically through C. quinoa, phenotypic variants inducing YM or green mild mosaic (MM) in tobacco were isolated. Aphid transmission, from different hosts, appears to be an effective approach for separating MM variants of CMV-G from YM variants. In particular, aphid transmission from zucchini proved to be very efficient in selecting for MM variants. In contrast, aphids transmitted only YM variants from tomato plants. Molecular characterization of CMV-G and its progeny resulted in their classification in the CMV subgroup IB, free of satellite RNA, being the first discovery of the subgroup IB in Greece. In the Solanaceae family (tobacco, tomato, pepper) YM variants induced more severe symptoms than the MM variants. YM and MM phenotype was stable in tobacco for all seven passages tried using the obtained YM and MM variants. Cross-protection experiments showed that an isolated MM variant was able to protect tobacco plants against a challenge infection by a YM variant.     

Mycoplasma infections in sheep, goats and yaks

Mycoplasmas were isolated from yak calves in Mongolia, the 1st experience worldwide with this species. They were also isolated from pathologically affected sheep and goat lambs. The disease was experimentally reproduced in yak calves as well as in sheep and goat lambs. Clinical manifestations were identical with those recordable from animals with spontaneous outbreak of the disease. This had been for the 1 time ever that a therapy was tested on animals with mycoplasmosis, using water with addition of oligodynamic silver. Something between 95 and 100% of all patients were clinically cured. The experimental vaccine involved was found to offer protection against mycoplasmosis to sheep and goat lambs. Damage association with mycoplasmosis was found to occur 1st in liver, spleen, and lymph nodes and to be subsequently proliferated to lung and other organs. Respiratory mycoplasmosis is the common definition used at present but is not in conformity with the pathological processes involved, as the major role is played by liver damage. Mycoplasmosis in these 3 species, therefore, should better be named mycoplasmosis bovi hepatica.     

Effect of virginiamycin supplementation on the performance of young grazing cattle

Three experiments have been conducted to study the effect of virginiamycin in young grazing-bulls (Experiment 1) and heifers (Experiments 2 and 3). Rotational grazing on permanent grassland was practised and animals were supplemented with 1 kg dried sugarbeet pulp per head daily. Control groups did not receive virginiamycin via the supplement, while it was incorporated at 120, 300 and 200 mg kg⁻¹ for the treatment groups in Experiments 1 to 3 respectively.
Average daily liveweight gain was significantly increased from 0·59 to 0·72 kg in Experiment 1 and from 0·73 to 0·81 kg in Experiment 3, but was not affected in Experiment 2 (0·51 vs 0·55 kg). Virginiamycin also increased liveweight gain per ha by 12, 7 and 13%, respectively, for the three experiments. Grazed area per animal was not enlarged to the same extent as the calculated daily energy allowances, which may suggest an improved digestibility and/or rumen fermentation.     

Equine infectious anemia in mules: virus isolation and pathogenicity studies

There appears to be a lack of information concerning responses of mules to natural infection or experimental inoculation with equine infectious anemia virus (EIAV). In the present study EIAV was isolated from mules, for the first time, and its pathogenicity in naturally infected and experimentally inoculated animals was investigated. Two naturally infected (A and B) and three EIAV free mules (C, D and E) were used for this purpose. Mule A developed clinical signs, whereas mule B remained asymptomatic until the end of the study. Mules C and D were each inoculated with 10ml of blood from mule A and developed signs of the disease; they were euthanatized or died at day 22 and 25 post-inoculation, respectively. Mule E served as a negative control. The virus was isolated from the plasma samples of mules with clinical signs of the disease (A, C and D), but not from the asymptomatic mule B. Both proviral DNA and viral RNA were amplified from blood and tissues of the infected animals by nested polymerase chain reaction (nPCR). Antibodies were not detected in the two experimentally infected mules until their natural death or euthanasia. Clinicopathological and laboratory findings showed that, in mules, EIAV produced clinical signs similar to those observed in horses and ponies. Nested PCR proved to be a rapid, sensitive and specific diagnostic method for the detection of EIAV, regardless of the disease stage.     

Highly diverse type of equine arteritis virus (EAV) from the semen of a South African donkey: short communication

Equine arteritis virus (EAV) was detected by RT-nested PCR in semen samples from a naturally infected South African donkey. Sequence analysis of the amplified ORF5 fragment revealed only 60 to 70% nucleotide identity to a panel of EAV reference sequences. The unique donkey EAV sequence was also found to be stable during passage in horses. The sequence data reported in this study indicate that the South African donkey variant might represent a new genotype of EAV. The distinct genetic properties of the South African asinine strain of EAV suggest a divergent evolution of this arterivirus in various host species or, alternatively, a possible role for African donkeys in the emergence of EAV in horses.     

Effect of therapeutic dietary biotin on the healing of uncomplicated sole ulcers in dairy cattle--a double blinded controlled study.

The objective of this study was to investigate the influence of orally administered biotin on the healing of uncomplicated sole ulcers in dairy cattle. In a double-blind controlled study, 24 dairy cows with a mild, uncomplicated sole ulcer on a lateral hind claw were given either 40 mg biotin per day or a placebo feed over a period of 50 days. An orthopaedic shoe was fitted to the medial claw of the affected foot. The healing process was assessed clinically and by histological examination of horn samples. In the biotin-treated animals, the newly formed epidermis covering the sole ulcers was found to be of significantly better histological quality after 50 days than at the start of the study. The significant improvement in histological horn quality found in the biotin-treated animals suggests that biotin exerts a positive influence on the healing of sole ulcers, however the study period of 50 days appears to have been too short to permit macroscopic detection of the improvement in horn quality.     

The effect of large doses of thiamine on the horse

Extract

In conjunction with a trial to determine the effect of nematode infestation upon the liveweight gain and wool production of young sheep (Brunsdon, unpublished), information was sought on the seasonal succession and availability on pasture of infective larvae of the principal trichostrongyle genera parasitic in sheep. Data were collected in the form of nematode populations found at postmortem examination of Romney lambs, previously maintained worm-free, which were exposed to risk of parasitism in the field. A series of these lambs ran with the flock in the above-mentioned trial and served as indicators of the abundance of infective larvae on the pasture.     

Effect of Flavonoids and Saponins Extracted from Black Bean (Phaseolus vulgaris L.) Seed Coats as Cholesterol Micelle Disruptors

Strategies for reducing plasma cholesterol have been focused on reducing the absorption or synthesis of cholesterol. The aim of this study was to correlate the content of flavonoids and saponins in black bean (Phaseolus vulgaris L.) seed coats extracts with a potential effect in lowering cholesterol absorption by the inhibition of cholesterol micellar solubility. Extracts with different flavonoids and saponins concentration were obtained from a Box-Behnken design used to optimize extraction temperature, stirring time and solvent composition. Major flavonoids and saponins were quantified by HPLC-PDA-ELSD and confirmed through mass spectrometry. Contrary to the flavonoid content, saponins were correlated to the inhibitory effect of cholesterol micelle solubility as an approach to a potential reduction of cholesterol absorption. Extracts with the highest saponin content strongly inhibited cholesterol micellization with values of 55.4?±?1.9 %, while stigmasterol used as control, only reached 12?±?2.3 % at the same concentration (5 mg/ml). The optimal extracting conditions for saponins were 25 °C, during 3 h in 85 % aqueous-methanol. Correlations of inhibitory effect of cholesterol micellar solubility with the content of each identified saponin suggested that the reduction of cholesterol micellization depends on the C-22 substitution of saponins.     

Pregnancy Rate in Zebu Cows with Two Different Postpartum Intervals Exposed to a Two-Bull Rotational System

In order to evaluate the reproductive effect of a rotational breeding system, the pregnancy rate of 69 Zebu cows was assessed, 32 late postpartum (211±93 days postpartum, LP) and 37 early postpartum (averaging 63±8 days after calving, EP). In the LP group, 60% (19/32) were cycling before exposure to the bulls but only 38% (14/37) in the EP group. The two groups were dissimilar from the start (p<0.05). The cows were exposed in a rotational breeding system, with six Brahman bulls with previous sexual experience paired off in three groups (A–B, C–D and E–F). The cows were exposed to each pair of bulls for a period of 3 weeks, with one week in between each period, when the females were without the presence of a male. In the LP group, the pair of bulls A–B obtained a pregnancy rate of 72% (23/32), whereas for bulls C–D the pregnancy rate was 33% (3/9) and for bulls E–F the pregnancy rate was 67% (4/6). In contrast, for the EP cows, bulls A–B obtained a pregnancy rate of 32% (12/37), bulls C–D a pregnancy rate of 67% (12/18) and for bulls E–F a pregnancy rate of 55% (6/11). The percentage pregnancy obtained using the pair of bulls A–B differed (p<0.05) between LP and EP, whereas for bulls C–D and E–F it was similar. The pregnancy rate was different (p<0.05) in the first 3 weeks of mating for the LP group, the highest number of cows becoming pregnant during the first 3 weeks of the study. In contrast, the highest number of pregnancies in EP occurred evenly during the last 6 weeks. This study suggests that the reproductive performance of pairs of bulls in a 9-week rotational programme with the overall pregnancy rate is similar (94% in the LP and 81% in the EP). However, the time taken for the females to become pregnant, and hence the performance of the bulls, is related to the average number of days postpartum for the cows.     

Effects of machine-milking on the bacterial flora of teat duct and mammary gland of ewes

In total, 308 paired-samples of teat duct material and milk, were collected before and 50-70 min after machine-milking, from 30 ewes. Samples were processed bacteriologically. For analysis of results, we compared changes in bacterial isolation following milking, for duct and milk samples; statistical significance was assessed by the Sign Test. Bacteria were isolated from 18 (6%) duct and 19 (6%) milk samples collected before the milking procedure; respective figures after it, were 81 (26%) and 33 (11%). In 77 (25%) cases, bacteriological findings in the two duct samples of each pair were different; in seven cases bacteria were isolated only before, whilst in 70 cases bacteria were isolated only after milking (P < 0.005); respective results for milk samples were 26 (8%): 6 and 20 cases (P = 0.693). The majority of bacterial isolates were staphylococci, accounting for 63% of 99 isolates. The milking procedure predisposes to entrance of bacteria into the teat duct; however, increased bacterial isolation from the teat did not result to increased mammary infections, likely as a consequence of defence mechanisms present in healthy teats.