Nasal immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis for improving ovarian responses and fertility in cross‐bred buffaloes

This study was conducted to determine the effect of immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis on ovarian responses and fertility in cross‐bred buffaloes. A total of 134 cross‐bred buffaloes were divided into four groups: groups T1 (n = 34), T2 (n = 35) and T3 (n = 31) were nasal immunized twice a day with 10 ml of 1 × 10¹⁰ CFU/ml of the C501 (pVAX‐asd‐IS) vaccine for 5, 3 and 1 day, respectively. Group C (n = 34) was nasal immunized with 10 ml PBS for 5 days. All animals were immunized twice with an interval of 14 days and administered with 200 μg of a GnRH analogue on day 28, 0.5 mg PGF_2α on day 35 and 200 μg of the same GnRH analogue on day 37. TAI was performed at 18 and 24 hr after the second GnRH treatment. Fourteen days after primary immunization, C501 (pVAX‐asd‐IS) elicited significant immune responses, and anti‐inhibin IgG antibody titres in group T1 were significantly higher (p < .01) than groups T3 and C. After the second GnRH treatment, the growth speed of the dominant follicles in group T1 was significantly faster (p < .05) than groups T3 and C. The number and diameter of large follicles (≥10 mm) as well as ovulatory follicles in group T1 were the greatest in all groups, resulting in a greater conception rate in buffaloes with positive anti‐inhibin antibodies. These results demonstrate that immunization with the C501 (pVAX‐asd‐IS) vaccine, coupled with the Ovsynch protocol, could be used as an alternative approach to improve reproductive performance in cross‐bred buffaloes.     

Scientific critique of the paper “Climatic distribution of citrus black spot caused by <Emphasis Type="Italic">Phyllosticta citricarpa</Emphasis>. A historical analysis of disease spread in South Africa” by Martínez-Minaya et al. (2015)

The global distribution of citrus black spot (CBS) disease, caused by Phyllosticta citricarpa, is climatically constrained, which is evident from its occurrence in citrus growing areas with warm, summer rainfall and its absence from areas with cooler, Mediterranean-type winter rainfall. Various epidemiological and modelling studies have supported this observation, predominantly estimating unsuitability for P. citricarpa in Mediterranean type climates, with no more than marginal suitability estimated at a few localities within some regions with Mediterranean type climates. The study by Martínez-Minaya et al. (European Journal of Plant Pathology, 143, 69–83, 2015), describes an historic sequence of recorded CBS occurrence in parts of South Africa, conducts an autocorrelation analysis and a correlative analysis with Köppen-Geiger climate zones and makes observations about the occurrence of certain Köppen-Geiger climate zones in the European Union. The study suggests that significant portions of the European Union and the broader Mediterranean basin are climatically similar to warm, summer rainfall areas in South Africa where P. citricarpa persists and causes CBS disease and concludes that the potential distribution of P. citricarpa is less constrained by climatic factors than spatial contagion. However, in this critique we expose methodological shortcomings in the Martínez-Minaya et al. (European Journal of Plant Pathology, 143, 69–83, 2015) study and conclude that the study grossly overestimated the extent of the geographical area that could support P. citricarpa, thereby rendering the findings scientifically unreliable.     

The use of adjuvants to improve spray deposition and Botrytis cinerea control on Chardonnay grapevine leaves

Spray adjuvants have the potential to improve deposition by effecting uniform distribution of the active ingredient on plant surfaces. In order to study whether such a qualitative improvement of spray deposition would lead to improved disease control, a laboratory experiment was conducted on artificially inoculated grapevine (cv. Chardonnay) leaves. Prior to inoculation with Botrytis cinerea conidia in a spore settling tower, leaves were sprayed to pre-runoff with 1 mL of a mixture of fenhexamid, a fluorescent pigment, and one of 15 selected commercial adjuvants to manipulate the deposition quality of a specific quantity of spray. Following an incubation period of 24 h at high relative humidity, leaf discs were plated onto Petri dishes with paraquat-amended water agar and rated for development of B. cinerea from isolated leaf discs 11 d later. Spray deposition on leaves was assessed with a spray assessment protocol using fluorometry, photomicrography and digital image analyses. B. cinerea incidences on the upper and lower surfaces of water-sprayed leaves averaged 90.4% and 95.8%, respectively. Despite full spray cover of leaves, applications with fenhexamid alone did not completely prevent infection and resulted in 34.6% and 40.8% B. cinerea incidences on the upper and lower surfaces of leaves, respectively. Through the addition of certain adjuvants, B. cinerea incidences were significantly reduced (incidences of 2.9–17.1% and 10.0–30.8%, respectively), while some adjuvants did not differ from the fungicide only treatment, even though they might have improved spray deposition. In a histopathology study using epifluorescence microscopy, distinct differences were observed in conidium mortality (20.5% vs. 31.2%), germination (60.4% vs. 51.4%) and germ tube lengths (27.8 μm vs. 19.7 μm) between Hydrosilicote and Solitaire in combination with fenhexamid, even though both adjuvants effected similar quantitative and qualitative spray deposition. The study clearly demonstrated the potential of adjuvants to improve the bio-efficacy of a fungicide directly through improved deposition on grapevine leaf surfaces, although bio-efficacy might be influenced by adjuvant mode of action in some cases.     

Photometric application of the MTT- and NRR-assays as biomarkers for the evaluation of cytotoxicity ex vivo in Eisenia andrei

In soils, the toxicity of noxious substances such as metals is determined by a number of different factors of chemical, physical, biological, and environmental origin. Addressing these factors as a sum of different parameters, we studied these effects in soil-dwelling indicator species by using ‘sub-cellular’ biomarkers, to see if they can provide an early stage response to environmental disturbances. ‘Sub-cellular’ biomarkers such as the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and the Neutral Red Retention (NRR) assays are well established for the assessment of cytotoxicity of toxicants on cell cultures. Both assays can be carried out in 96-well microplates and evaluated photometrically. To be able to assess the risk potential of contaminated soils using these assays, without changing the soils chemically or physically to accommodate cell cultures, the assays were adapted for ex vivo use. The assays were performed on coelomic liquid extracted from exposed earthworms. Eisenia andrei was exposed in a standardized laboratory soil substrate (OECD soil) spiked with cadmium at a range of concentrations known to be cytotoxic to earthworms. Earthworms were also exposed to ultramafic soils as field-relevant soil substrates, containing high concentrations of a mixture of metals of natural origin such as chromium, cobalt, manganese, and nickel. A significant response was measured with both assays in the coelomocytes of the earthworms exposed to the Cd-spiked OECD soil as well as to the ultramafic soils. It was shown that as biomarkers for the assessment of ‘sub-cellular’ toxicity, the photometrical application of the MTT and the NRR assay promised to be rapid, objective, and easily conductible tools.     

Bragg diffraction from crystallized ion plasmas

Single crystals of a one-component plasma were observed by optical Bragg diffraction. The plasmas contained 10(5) to 10(6) single-positive beryllium-9 ions (9Be+) at particle densities of 10(8) to 10(9) per cubic centimeter. In approximately spherical plasmas, single body-centered cubic (bcc) crystals or, in some cases, two or more bcc crystals having fixed orientations with respect to each other were observed. In some oblate plasmas, a mixture of bcc and face-centered cubic ordering was seen. Knowledge of the properties of one-component plasma crystals is required for models of white dwarfs and neutron stars, which are believed to contain matter in that form.     

Hydrogen radicals, nitrogen radicals, and the production of O3 in the upper troposphere

The concentrations of the hydrogen radicals OH and HO2 in the middle and upper troposphere were measured simultaneously with those of NO, O3, CO, H2O, CH4, non-methane hydrocarbons, and with the ultraviolet and visible radiation field. The data allow a direct examination of the processes that produce O3 in this region of the atmosphere. Comparison of the measured concentrations of OH and HO2 with calculations based on their production from water vapor, ozone, and methane demonstrate that these sources are insufficient to explain the observed radical concentrations in the upper troposphere. The photolysis of carbonyl and peroxide compounds transported to this region from the lower troposphere may provide the source of HOx required to sustain the measured abundances of these radical species. The mechanism by which NO affects the production of O3 is also illustrated by the measurements. In the upper tropospheric air masses sampled, the production rate for ozone (determined from the measured concentrations of HO2 and NO) is calculated to be about 1 part per billion by volume each day. This production rate is faster than previously thought and implies that anthropogenic activities that add NO to the upper troposphere, such as biomass burning and aviation, will lead to production of more O3 than expected.     

Atmospheric residence time of CH3Br estimated from the junge spatial variability relation

The atmospheric residence time for methyl bromide (CH3Br) has been estimated as 0.8 +/- 0.1 years from its empirical spatial variability relative to C2H6, C2Cl4, CHCl3, and CH3Cl. This evaluation of the atmospheric residence time, based on Junge's 1963 general proposal, provides an estimate for CH3Br that is independent of source and sink estimates. Methyl bromide from combined natural and anthropogenic sources furnishes about half of the bromine that enters the stratosphere, where it plays an important role in ozone destruction. This residence time is consistent with the 0.7-year value recently calculated for CH3Br from the combined strength estimates for its known significant sinks.     

Comparison of the Johne''s absorbed EIA and the complement-fixation test for the diagnosis of Johne''s disease in cattle

A commercially available absorbed ELISA for the diagnosis of Johne's disease (JD) (paratuberculosis) in cattle, the Johne's Absorbed EIA, was compared with the conventional complement-fixation test (CFT) used in Australia. Stored plasma from 3 Victorian dairy herds with a history of JD, sera from specimens submitted from animals showing clinical signs of JD and sera from the US National Repository for Paratuberculosis Specimens were used to determine the sensitivity of each test. The EIA detected 48.8% of 43 Australian animals with subclinical JD, while the CFT detected only 12 (21.4%) of 56 subclinically affected cattle. Of 150 subclinically infected US cattle, the EIA detected 47.3% and the CFT detected 52.0%. The EIA detected 59.7% of animals which at the time of sampling were shedding Mycobacterium paratuberculosis in their faeces, but showed no clinical signs of JD, while the CFT detected 57.3%. The EIA correctly identified 88.2% of 136 histologically confirmed clinical cases, and the CFT detected 83.4%. The specificity of each test was determined by testing sera collected at slaughter from animals residing in a known JD-free area of Australia, and from samples from the US National Repository of Paratuberculosis Specimens collected from certified-free herds in Wisconsin. The EIA was found to have a specificity of 99.8% when 998 Australian animals were used as the test population, and 99.0% when 196 US animals were used. The specificity of the CFT using Australian samples was 96.9% and 95.2% using American samples.     

Seasonal incidence of Karoo paralysis in relation to the infestation density of female Ixodes rubicundus

The epidemiology of Karoo paralysis, induced by the tick Ixodes rubicundus, was investigated in a paralysis-enzootic area of the southwestern Orange Free State, Republic of South Africa. Sheep became paralysed during the last week of April and the first week of May during the 1985 and 1986 seasons, respectively. Paralysis cases were recorded up until July, but most paralysis cases occurred during the first week of May with a second but smaller peak occurring during June. Tick numbers on paralysed sheep closely followed peak periods in paralysis. A total of 65 and 50% of the naive sheep became paralysed during the 1985 and 1986 seasons, respectively. The seasonal pattern of tick infestation by female I. rubicundus was similar on both previously exposed and on naive sheep. The results of this study indicate that a certain number of ticks per kilogram host body mass is required before symptoms of paresis of paralysis are observed. In general the tick burden per kilogram host mass was significantly greater in paralysed compared to non-paralysed sheep. The range in the number of ticks per kilogram host mass may, however, overlap between non-paralysed and paralysed groups. Previously exposed or previously paralysed sheep are less susceptible to paralysis in the following year. Differences in susceptibility between individual sheep seems likely.     

A field outbreak in Ile-de-France sheep of a cardiotoxicosis caused by the plant Pachystigma pygmaeum (Schltr) Robyns (Rubiaceae)

A field outbreak of Pachystigma pygmaeum intoxication in sheep is described. Noteworthy clinical signs were: respiratory distress, apathy and subcutaneous oedema of mainly the head. Gross changes included cardiomegaly, centrilobular hepatic necrosis and effusion of body cavities. Microscopically myocardial fibrosis, affecting predominantly the endocardium of the apex, left free ventricular wall and interventricular septum, was most striking in the majority of animals, whilst myofibre atrophy was present in 1 sheep.