An Investigation of the Ability of the Glutaraldehyde Test to Distinguish between Acute and Chronic Inflammatory Disease in Horses

The Glutaraldehyde test (GT), a rapid and inexpensive test, has been utilized empirically for many years in bovine practice for diagnosing inflammatory diseases. GT is used primarily to demonstrate increased serum concentrations of fibrinogen and globulin. Glutaraldehyde binds with free amino groups in fibrinogen and immunoglobulin to create a clot in a first degree chemical reaction. The clotting time of the GT estimates the content of proteins produced in response to inflammation. The applicability of GT for diagnosing inflammation in the horse has never been investigated. The objective of this study was to determine the ability of GT to distinguish between acute and chronic inflammatory disease in horses. Thirty-seven horses with suspected inflammatory diseases were evaluated using the GT, history, complete clinical examination and routine blood analysis. GT-times, laboratory results and clinical outcome were compared statistically. Horses that were determined to be acutely affected (based on history, clinical examination and routine blood analysis) tended to have a negative GT (75%). Results of the GT did not correlate with blood fibrinogen concentration. Positive GT also predicted a fatal outcome in 69% of the clinical cases. The results of this trial indicate that GT can be a useful screening test to distinguish between acute and chronic inflammatory disease in horses.     

Ovum Pick-up in Sheep: a Comparison between Different Aspiration Devices for Optimal Oocyte Retrieval

In vivo ovum pick-up (OPU) in sheep may be improved with a proper choice of aspiration elements (needle and tubing) and aspiration vacuum pressure. In the present study, two experiments were carried out. In Expt 1, visible follicles in ovaries of slaughtered ewes (treated separately according to their diameters: small<3 mm, medium 3-5 mm and large>5 mm) were aspirated using different combinations of the three studied factors such as aspiration flow rate (10, 20, 30, 40 and 50 ml water/min), needle gauge (18 and 20 G) and tubing inner diameter (1, 2 or 3 mm internal diameter). In Expt 2, a study with two 18 G needles of different lengths (18 G: 82 mm; 18 GL: 600 mm) was carried out, using ovaries obtained post-mortem, and performing in vivo laparoscopic follicular aspiration on ewes. We considered good quality oocytes as those with both complete compact cumulus and a homogeneous cytoplasm. Recovery rate, proportion of good quality oocytes (good quality oocytes/100 oocytes recovered) and overall efficiency (good quality oocytes/100 follicles aspirated) were noted. In Expt 1, aspiration flow rate affect remarkable proportion of good quality oocytes (69.5%, 50.5%, 44.8%, 36.5% and 28.3% for flows from 10 to 50 ml/min respectively, p<0.05). Needle gauge did not affect aspiration device efficiency. Thin and intermediate tubings were more effective (overall efficiency rates: 34.9%, 32.3% and 28.1% for 1, 2 and 3 mm respectively, p<0.05). Follicle size did not affect recovery rate, but proportion of good quality oocytes was higher for large (77.9%) and medium (64.4%) follicles (p<0.05). Finally, some combinations of the aspiration device showed greater effectiveness. In Expt 2, needle length did not influence recovery rate, but good quality oocytes rate was significantly modified both post-mortem and in vivo (good quality rate for 18 G vs 18 GL needles: 69.5% vs 47.7% and 58.1% vs 25.4%, post-mortem and in vivo respectively, p<0.05). We conclude that low-aspiration flow rates (10 and 20 ml/min) with thin or intermediate tubings (1 and 2 mm), and any short needle (18 G or 20 G) are the most adequate aspiration factors for OPU in sheep.     

Growth response of six weed species and spring barley (Hordeum vulgare) to increasing levels of nitrogen and phosphorus

A glasshouse experiment was carried out to investigate the influence of increasing levels of nitrogen and phosphorus on the growth of six common weed species growing alone or in competition with spring barley (Hordeum vulgare). Capsella bursa‐pastoris, Chenopodium album, Papaver rhoeas, Sinapis arvensis, Spergula arvensis, Viola arvensis and spring barley were grown in pots with different levels of nitrogen (0, 30, 60, 90, 120 and 150 kg N ha^?1) or phosphorus (0, 10, 20, 30, 40 and 60 kg P ha^?1). The aboveground parts of the plants were harvested after 7 weeks and the dry weight of shoots, percentage N and P content of the shoot and uptake of N and P were determined. A linear or a polynomial model was used to describe the data. Growing alone, Spergula arvensis was the only weed species that increased its dry weight at the same rate as barley. Weed species with low dry weight increase had larger increases in percentage N or P content than barley, indicating a luxury accumulation of nutrients. The uptake of N and P per pot did not differ much between weeds and barley. V. arvensis and P. rhoeas accumulated least nutrients (per cent of dry matter) and Spergula arvensis accumulated most. Weeds grew poorly in competition with barley. The percentage N and P content in barley did not change when they grew in competition with weeds.     

Characterization of Pyricularia grisea in the United States Using Independent Genetic and Molecular Markers

ABSTRACT A total of 540 isolates of Pyricularia grisea from rice in the United States were examined for vegetative compatibility, MGR586 DNA fingerprint diversity, and mating type based on hybridization with the mat1-1 and mat1-2 sexual mating type alleles. The collections contained both archived and contemporary field isolates representative of the known MGR586 lineages and races that occur throughout the United States. Complementary nitrate nonutilizing (nit) or sulfate nonutilizing (sul) mutants were used to assess vegetative compatibility in P. grisea. There was a complete correspondence between vegetative compatibility groups (VCGs), MGR586 lineage, and mating type among 527 contemporary isolates (collected between 1991 and 1997) from Arkansas, Louisiana, Missouri, Mississippi, and Texas; all isolates in MGR586 lineages A, B, C, and D belonged to VCGs US-01, US-02, US-03, and US-04, respectively. In addition, all isolates tested in VCGs US-01 and US-04 had the mat1-1 mating type allele whereas those in VCGs US-02 and US-03 had the mat1-2 allele. The strict association of independent markers during this sample period was consistent with a strictly asexual mode of reproduction. However, examination of archived isolates collected in the 1970s and 1980s and contemporary isolates revealed an incongruent relationship between the independent markers. MGR586 C and E isolates were vegetatively compatible which indicated that multiple robust MGR586 delineated lineages could be nested within certain VCGs. Although isolates in lineages C and E were vegetatively compatible, they were of opposite mating type. Several hypotheses, including recombination, could account for the incongruence between the various markers. Among the eight MGR586 lineages (A through H) that occur in the United States, all isolates in lineages A, D, E, G, and H had the mat1-1 allele, whereas isolates in lineages B, C, and F had the mat1-2 allele. Nit mutants can be recovered relatively easy from P. grisea and should allow large numbers of individuals within a population to be assessed for vegetative compatibility. VCGs may prove to be an effective multilocus marker in P. grisea. Thus, VCGs should be a useful means for characterizing genetic structure in populations of the rice blast fungus worldwide, provide a useful genetic framework to assist in interpreting molecular population data, and may provide insight into potential sexual or asexual recombination events.     

Investigation of bovine haemoplasmas and their association with anaemia in New Zealand cattle

CASE HISTORY AND CLINICAL FINDINGS: A dairy cow, from a herd in the Waikato region of New Zealand, was reported with regenerative anaemia on 12 September 2014. Testing of blood from the animal using PCR assays for Theileria orientalis produced a negative result for both Chitose and Ikeda types.

LABORATORY FINDINGS: Using PCR and DNA sequencing, blood from the cow was positive for Candidatus Mycoplasma haemobos. Further testing of another 12 animals from the case herd, 27 days after the affected cow was first reported, showed 11 animals were positive for Candidatus M. haemobos or Mycoplasma wenyonii in the PCR. None of these cattle were clinically anaemic or positive for T. orientalis Ikeda type using PCR.

A convenience sample of 47 blood samples from cattle throughout New Zealand, submitted to the Investigation and Diagnostic Centre (Ministry for Primary Industries) for surveillance testing for T. orientalis Ikeda, was selected for further testing for bovine haemoplasmas. Of these samples, 6/47 (13%) and 13/47(28%) were positive for M. wenyonii and Candidatus M. haemobos, respectively. There was no difference in the proportion of samples positive for the bovine haemaplasmas between cattle with anaemia that were negative for T. orientalis (6/20, 33%), or without anaemia or T. orientalis (10/18, 56%), or from cattle herds experiencing anaemia and infection with T. orientalis Ikeda type (3/9, 33%).

DIAGNOSIS: Bovine haemoplasmosis.

CLINICAL RELEVANCE: The presence of bovine haemoplasmas in blood does not establish causality for anaemia in cattle. Diagnosis of anaemia associated with haemoplasmosis would require exclusion of other causes of regenerative anaemia and an association of the agent with anaemia in affected cattle herds. The data collected in this study did not provide evidence that bovine haemoplasmas were associated with a large number of outbreaks of anaemia in cattle in New Zealand.     

Resynchronising returns to service in anoestrous dairy cows in the South Island of New Zealand

AIMS: To investigate the effect of targeted resynchronisation of cows treated for non-observed oestrus before the planned start of mating (PSM), that were not detected in oestrus or pregnant 23 days after treatment (phantom cows), on the proportion pregnant at 42 days after PSM and the end of mating.

METHODS: Farm staff from eight herds in two regions of the South Island of New Zealand identified 1,819 cows not showing oestrus by 10 days before PSM. These cows were treated with intravaginal progesterone for 7 days, and I/M gonadorelin 10 days and 1 day before PSM. Three days before PSM they were injected with cloprostenol and equine chorionic gonadotrophin, with fixed time artificial insemination (FTAI) at PSM. By 23 days after PSM, 1,218 cows had not returned to oestrus. Of these, 161 cows confirmed not pregnant by transrectal ultrasonography were randomly assigned to no treatment (control group; n=74) or were resynchronised 25 days after PSM using the same treatment programme as above, with FTAI 35 days after PSM (n=87). All cows that returned to oestrus were artificially inseminated until 42 days after PSM, when natural mating was used. All cows were examined using transrectal ultrasonography 80 to 90 days after PSM to confirm conception dates.

RESULTS: Of the 1,819 anoestrous cows treated before PSM, 526 (29 (95% CI=23.1–34.0)%) had not been observed in oestrus by 23 days after PSM and had not conceived, so were diagnosed as phantoms cows. For resynchronised cows, 42/87 (48 (95% CI=37.8–58.8)%) were pregnant by 42 days after PSM compared to 21/74 (28 (95% CI=18.1–38.7)%) control cows (p=0.009). At the end of mating 58/87 (67 (95% CI=56.6–76.7)%) cows in the resynchronised group were pregnant and 46/74 (62 (95% CI=50.9–73.2)%) in the control group (p=0.554). The hazard of conception from 21 to 42 days after PSM was 1.9 (95% CI=1.07–3.12) times greater for resynchronised than control cows (p=0.026).

CONCLUSION: In cows not observed in oestrus and treated before PSM, resynchronisation increased the proportion pregnant by 42 days after PSM.

CLINICAL RELEVANCE: The benefit of resynchronisation depends on the number of anoestrous cows before PSM and the number of phantom cows after PSM. However at the herd-level it is likely that providing advice to reduce the known risk factors for cows not being observed in oestrus before the PSM may well be more cost effective than identifying and treating a sub-population of phantom cows.     

Vertical Flux of Biogenic Carbon in the Ocean: Is There Food Web Control?

Models of biogenic carbon (BC) flux assume that short herbivorous food chains lead to high export, whereas complex microbial or omnivorous food webs lead to recycling and low export, and that export of BC from the euphotic zone equals new production (NP). In the Gulf of St. Lawrence, particulate organic carbon fluxes were similar during the spring phytoplankton bloom, when herbivory dominated, and during nonbloom conditions, when microbial and omnivorous food webs dominated. In contrast, NP was 1.2 to 161 times greater during the bloom than after it. Thus, neither food web structure nor NP can predict the magnitude or patterns of BC export, particularly on time scales over which the ocean is in nonequilibrium conditions.