Detection of hypoglycin A in the seeds of sycamore (Acer pseudoplatanus) and box elder (A. negundo) in New Zealand; the toxin associated with cases of equine atypical myopathy

CASE HISTORY AND CLINICAL FINDINGS: During April and May 2014 four horses aged between 5 months and 9 years, located in the Canterbury, Marlborough and Southland regions, presented with a variety of clinical signs including recumbency, stiffness, lethargy, dehydration, depression, and myoglobinuria suggestive of acute muscle damage. Two horses were subjected to euthanasia and two recovered. In all cases seeds of sycamore maple (Acer pseudoplatanus) or box elder (A. negundo) were present in the area where the horse had been grazing.

LABORATORY INVESTIGATION: The samaras (seeds) of some Acer spp. may contain hypoglycin A, that has been associated with cases of atypical myopathy in Europe and North America. To determine if hypoglycin A is present in the samaras of Acer spp. in New Zealand, samples were collected from trees throughout the country that were associated with historical and/or current cases of atypical myopathy, and analysed for hypoglycin A. Serum samples from the four cases and four unaffected horses were analysed for the presence of hypoglycin A, profiles of acylcarnitines (the definitive diagnosis for atypical myopathy) and activities of creatine kinase and aspartate aminotransferase.Markedly elevated serum activities of creatine kinase and aspartate aminotransferase, and increased concentrations of selected acylcarnitines were found in the case horses. Hypoglycin A was detected in the serum of those horses but not in the healthy controls. Hypoglycin A was detected in 10/15 samples of samaras from sycamore maple and box elder from throughout New Zealand.

DIAGNOSIS: Cases of atypical myopathy were diagnosed on properties where samaras containing hypoglycin A were also found.

CLINICAL RELEVANCE: Sycamore and box elder trees in New Zealand are a source of hypoglycin A associated with the development of atypical myopathy. If pastured horses present with clinical and biochemical signs of severe muscle damage then the environment should be checked for the presence of these trees. Horses should be prevented from grazing samaras from Acer spp. in the autumn.     

The first reported outbreak of equine herpesvirus myeloencephalopathy in New Zealand

CASE HISTORY AND CLINICAL FINDINGS: On 9 January 2014 (Day 0) a mare from a stud farm in the Waikato region presented with urinary incontinence without pyrexia. Over the following 33 days 15 mares were clinically affected with neurological signs. All but one mare had a foal at foot. The most commonly observed clinical signs were hind limb paresis and ataxia. In some cases recumbency occurred very early in the course of disease and seven mares were subject to euthanasia for humane reasons.

LABORATORY FINDINGS: Equid herpesvirus (EHV) type 1 was detected using PCR in various tissues collected post mortem from two mares with neurological signs. DNA sequencing data from the DNA polymerase gene of the virus showed a nucleotide transition at position 2254, a mutation encoding amino acid D₇₅₂ that is highly associated with the neuropathogenic genotype of EHV-1. In total 12/15 mares were confirmed positive for EHV-1 on PCR. Results from a virus neutralisation test and ELISA on paired serum samples, and PCR on whole blood and nasal swabs, indicated that of four paddocks in a high-risk area where a cluster of cases had occurred, 20/21 (95%) horses were likely to have been exposed or were confirmed infected with EHV-1. Subsequent to the outbreak two mares aborted, one at 9 months and one at 10 months of gestation. The cause of abortion was confirmed as EHV-1 with the same genotype as that involved in the outbreak.

DIAGNOSIS: Equine herpesvirus myeloencephalopathy.

CLINICAL RELEVANCE: The outbreak described shows the considerable impact that can occur in outbreaks of equine herpesvirus myeloencephalopathy in New Zealand. Early biosecurity controls not only reduced the effect on the farm but mitigated the potential for the virus to spread to other horse enterprises.     

Supplementation of Slow‐Release Melatonin Improves Recovery of Ovarian Cyclicity and Conception in Summer Anoestrous Buffaloes (Bubalus bubalis)

The role of melatonin as a protective neurohormone against restoring cyclicity in summer anoestrous animals in photoperiod species has gained wider acceptance. This study was designed to uncover the evidence the slow‐release melatonin (MLT) has on initiation of ovarian cyclicity and conception rate (CR) in summer anoestrous buffaloes. Thus, buffaloes diagnosed as summer anoestrous (absence of overt signs of oestrus, concurrent rectal examination and radioimmunoassay for serum progesterone at 10 days interval) were grouped as untreated (Group I, sterilized corn oil, n = 8) and treated (Group II, single subcutaneous injection of MLT @18 mg/50 kg bwt in sterilized corn oil, n = 20). Animals treated and detected in oestrus were artificially inseminated (AI) followed by division into Group III (second dose of MLT on 5th day post‐AI, n = 8) and Group IV (no melatonin administration, n = 10). Blood samples were collected at 4 days interval for estimation of serum MLT, progesterone and oestrogen using radioimmunoassay kit. Mean oestrous induction rate (OIR), oestrous induction interval (OII), interoestrous interval (IOI) and CR were estimated. Compared to control, concentration of melatonin was significantly (p < 0.05) higher in treated group ranging from 14.34 ± 1.72 to 412.31 ± 14.47 pg/ml whereas other two hormones did not show any concentration difference. Melatonin‐administered buffaloes showed significantly (p < 0.05) higher (90%) OIR with OII of 18.06 ± 1.57 days. Results showed improvement in conception rate in buffaloes administered with post‐insemination melatonin. It can be concluded from the study that slow‐release melatonin supplementation restored cyclicity in summer anoestrous animals resulting in improvement in conception rate in buffaloes.     

Genetic Diversity and Yield Stability of Potato(Solanum tuberosum L.)

The present study was under taken to investigate the genetic distance and the performance of thirty one potato genotypes in diverse environments.Multivariate and canonical analysis were used to study genetic diversity among the genotypes and stability parameters were estimated according to Eberhart and Russell,Parkinge and Jinks and Freeman and Perkins models for plant height,branch number per plant,tuber number and tuber weight.Thirty one genotypes were grouped in to six clusters with the help of Mahalonobis distance and canonical analysis.The stability analysis revealed that yield was greatly influenced by the environments.Most of the studied genotypes were sensitive for yield with the environment.For over-all performance 'Hera','Chamak','Patnai','Lala pakri','TPS-7','TPS-364','Somerset' and 'Superior' were the best genotypes for tuber yield and breeding material for potato breeding.     

Endophytic colonization of white jute (Corchorus capsularis) plants by different Beauveria bassiana strains for managing stem weevil (Apion corchori)

Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales), an entomopathogenic fungus, was introduced through seed inoculation with spore suspension as an endophyte in white jute (Corchorus capsularis L.), a bast fiber crop. Out of nine B. bassiana strains, seven, viz., ITCC 6063, ITCC 4512, ITCC 4563, ITCC 5562, ITCC 4796, ITCC 5408 and ITCC 4705, became established as endophytes. The endophytic colonization was initially detected by cultivation on selective medium. Colonization was confirmed by polymerase chain reaction (PCR) using sequence characterized amplified region (SCAR) marker. Endophytic B. bassiana strains colonized in leaves of all the plants grown from treated seeds. However, the colonization frequency varied among the strains. The highest colonization frequency (70.09%) was recorded in ITCC 6063 followed by ITCC 5562 (67.67%) and ITCC 5408 (64.17%); ITCC 4512 exhibited the lowest (42.54%) colonization. ITCC 4925 and ITCC 4644 did not show any colonization. Endophytic colonization of B. bassiana reduced stem weevil infestation under pot culture. ITCC 5408 and ITCC 6063 were found most efficient, with only 10.44% and 14.06% infested plants, respectively.     

Co-inoculation of potassium solubilizing and nitrogen fixing bacteria on solubilization of waste mica and their effect on growth promotion and nutrient acquisition by a forage crop

Waste mica, a potassium-bearing mineral, is a by-product of mica industry; however, its potassium (K)-supplying capacity for crop production is not well understood. A greenhouse trial was made to study the effect of co-inoculation of potassium solubilizing (Bacillus mucilaginosus) and nitrogen (N) fixing (Azotobacter chroococcum A-41) bacteria on solubilization of waste mica (a potassium-bearing mineral) and their effects on growth promotion and nutrient uptake by a forage crop of sudan grass (Sorghum vulgare Pers.) in a Typic Haplustalf. Results revealed that significantly higher biomass accumulation and nutrient acquisition were obtained in all the pots treated with mica and/or bacterial strain as compared to control. Data indicated that co-inoculation of waste mica with B. mucilaginosus and A. chroococcum A-41 resulted in highest biomass production and nutrient acquisition. Co-inoculation of bacterial strains maintained consistently highest amounts of available K and N in soils even at 150 days of crop growth than other treatments. B. mucilaginosus strain was more effective and potent K solubilizer than A. chroococcum A-41. Thus, co-inoculation of potassium solubilizing and nitrogen fixing bacteria to waste mica could be a promising and alternative option for utilizing this potent source as K fertilizer to crops and maintaining greater nutrients availability in soil. Further studies are necessary to see the effects of these bacterial strains on mobilization of potassium-bearing minerals under field conditions.     

Evidence of mixed infection of peste des petits ruminants virus and bluetongue virus in a flock of goats as confirmed by detection of antigen, antibody and nucleic acid of both the viruses

A mixed infection with peste des petits ruminants virus (PPRV) and bluetongue virus (BTV) occurred in goats which exhibited symptoms characteristic of PPR. A number of samples were collected from ailing or dead goats for labrotory diagnosis. Antibody to BTV and PPRV was detected in sera samples by competitive ELISA. No PPRV antigen was detected in tissue samples like lung and spleen, however, presence of PPRV antigen in some sera samples was confirmed by sandwich ELISA. All the blood samples collected from the ailing animals were found positive for BTV antigen by a sandwich ELISA. BTV- and PPRV nucleic acids were amplified from the pooled blood and tissue samples respectively by RT-PCR assays. The identity of the amplicons was confirmed by cloning and sequencing. All these tests confirm that the goats were infected with PPRV and BTV simultaneously. Isolation of viruses from the clinical samples is underway.     

Comparative analysis of genetic diversity in Citrus germplasm collection using AFLP,SSAP, SAMPL and SSR markers

In this study we evaluate the informativeness and efficiency of Amplified Fragment Length Polymorphism (AFLP), Sequence-Specific Amplified Polymorphism (S-SAP), Selectively Amplified Microsatellite Polymorphic Loci (SAMPL) and Simple Sequence Repeat (SSR) markers for genetic diversity, phylogenetic relationship among the Citrus species and mapping ability of the marker system. The SSR exhibited relatively higher level of polymorphism information content in terms of the expected heterozygosity, than that of the AFLPs, SSAPs and SAMPLs. For each marker system, average level of the discriminating potential was very close to the actual discriminating potential. Similarity matrices showed weak, yet significant correlations when Mantel's test was applied. The highest positive (0.72) correlation was found between the AFLP and SSAP markers. The SSR and SAMPL markers were poorly correlated. The dendrogram topology among the four marker systems had high similarity. Taken together, the SSAP and SAMPL were highly efficient in detecting genetic similarity in Citrus, while the SSR may be more useful for segregation studies and genome mapping in Citrus. The SSAP and SAMPL markers could be useful for Citrus genome mapping in combination with AFLP and SSR markers. To our knowledge, this was the first detail report of a comparison of performances among AFLP, SSR and retrotrasposon based molecular marker technique on a set of samples of Citrus. Our result provides guidance for future efficient use of these molecular methods in genetic analysis of Citrus sp. and its relatives.     

Senescence of Flag Leaf and Glume in Rice: Role of Grains during Source-Sink Modification by Physical and Chemical Means

Senescence of flag leaf and glume of rice ( Oryza sativa L. cv. Ratna ) at different source sink ratios, through manipulating the plants by physical and chemical means, were analyzed. As judged from the chlorophyll retention capacity, induction of sterility deferred senescence of flag leaf and glume at both grain filling and post grain maturation period. Senescence of the glumes was promoted in the plants whose panicles were darkened. The time needed for achieving maximum 1000 grain dry weight was quickened, the harvest index (HI) was lowered and the flag leaf senescence was delayed by such manipulation. Spraying of urea deferred senescence of the flag leaf as well as increased 1000 grain dry weight and HI. The time taken for achieving maximum 1000 grain dry weight was, however, delayed. Senescence of glumes was promoted by α-HPMS spray, while the same was delayed by PMA spray on the whole plant. Both the chemicals deferred flag leaf senescence and decreased 1000 grain dry weight and HI. Senescence of the flag leaf at post grain filling period was also delayed by the above physical and chemical manipulations compared to that of control, the highest delaying effect was recorded in the flag leaf of sterile plant.
It is concluded that nutrient drainage is the primary cause of nonsequential senescence of the flag leaf of rice (cv. Ratna ). The involvement of senescence signal if any, is perhaps of secondary importance.